Use of lipid conjugates in the treatment of disease

ABSTRACT

This invention provides a method of preventing or treating a dermatologic condition including, inter alia, psoriasis, contact dermatitis, and atopic dermatitis, in a subject, the method includes the step of administering to said subject a compound comprising a lipid or phospholipid moiety bound to a physiologically acceptable monomer, dimer, oligomer, or polymer, and/or a pharmaceutically acceptable salt or a pharmaceutical product thereof, in an amount effective to treat the subject suffering from a dermatologic condition.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional application of U.S. patent applicationSer. No. 11/524,519, filed Sep. 21, 2006, which is acontinuation-in-part application of U.S. patent application Ser. No.10/989,607, filed Nov. 17, 2004 and U.S. patent application Ser. No.10/952,496, filed Sep. 29, 2004, which are continuation-in-partapplications of U.S. patent application Ser. No. 09/756,765, filed Jan.10, 2001, now U.S. Pat. No. 7,034,006 which claims priority from U.S.Provisional Patent Application Ser. No. 60/174,907, filed Jan. 10, 2000,which are all hereby incorporated by reference.

FIELD OF THE INVENTION

This invention provides compounds and methods of use thereof insuppressing, inhibiting, preventing, or treating a dermatologiccondition in a subject, including, inter alia, psoriasis, contactdermatitis, atopic dermatitis, and seborrheic dermatitis.

BACKGROUND OF THE INVENTION

Lipid-conjugates are thought to inhibit the enzyme phospholipase A2(PLA2, EC 3.1.1.4). Phospholipase A2 catalyzes the breakdown ofphospholipids at the sn-2 position to produce a fatty acid and alysophospholipid. The activity of this enzyme has been correlated withvarious cell functions, particularly with the production of lipidmediators such as eicosanoid production (prostaglandins, thromboxanesand leukotrienes), platelet activating factor and lysophospholipids.Lipid-conjugates may offer a wider scope of protection of cells andorganisms from injurious agents and pathogenic processes, including theprevention and treatment of dermatologic conditions.

Dermatologic conditions are far more prevalent than once thought andcost an estimated $37.17 billion annually in the US, according to acomprehensive study jointly released in April 2005 by the AmericanAcademy of Dermatology Association and the Society for InvestigativeDermatology. These estimates consider the costs of doctor visits,over-the-counter and prescription medicine costs and lost workproductivity. The new study found that at any given time, one in everythree people in the U.S. suffers from a skin disease—exceeding theprevalence of obesity, hypertension and cancer. Among the mosteconomically burdensome skin disorders are skin ulcers and wounds,melanoma, nonmelanoma skin cancer, atopic dermatitis and acne, the costof which totaled $22.46 billion in 2004. It is estimated that NIHresearch expenditures on skin diseases will total nearly $172 million in2005.

Dermatitis, which is sometimes called eczema, is an inflammationreaction of the skin to various external and internal causes and is themost common among the skin diseases. Typical clinical features in acutestage dermatitis include swelling erythema, followed by the formation ofpapules and serous papules on the erythema. After the formation ofvesicles in the skin, pustules form, followed by the erosion, crustingand desquamation of the skin. Only then does the skin begin to heal.When dermatitis turns chronic, thickening, lichenification andpigmentation of the skin all result and, in most cases, accompanied byitching. Histologically, dermatitis is characterized by swelling amongepidermal cells (in a spongy state) during the acute stage. Contactdermatitis, atopic dermatitis, seborrheic dermatitis, etc. are includedamong recognized categories of dermatitis.

External application of a steroidal agent (ointment or the like) is themost effective therapy to date and no therapeutic method to replace ithas yet been established. However, steroid preparations also cause agreat variety of adverse reactions. There have been various reports onthe side effects of steroid preparations and, in the case of agents forexternal use such as ointments, direct harmful effects such as thethinning, shrinking and flushing of the skin. The severe adversereactions caused by steroid preparations used as remedies fordermatologic conditions have led to a demand in the medical field forsafer pharmaceuticals which have fewer side effects.

SUMMARY OF THE INVENTION

In one embodiment, the invention provides a method of preventing adermatologic condition in a subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof.

In another embodiment, the invention provides a method of treating adermatologic condition in a human subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000;        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof, wherein X is hyaluronic acid and L is        phosphatidylethanolamine.

In one embodiment, X in general formula (A) represents a polysaccharide.In one embodiment, the polysaccharide is carboxymethylcellulose, inanother embodiment, the polysaccharide is dextran, while in anotherembodiment, the polysaccharide is a glycosaminoglycan. In oneembodiment, the glycosaminoglycan is hyaluronic acid, in anotherembodiment, the glycosaminoglycan is chondroitin sulfate, while inanother embodiment, the glycosaminoglycan is heparin. In one embodiment,L in general formula (A) is phosphatidylethanolamine, which in oneembodiment is dipalmitoyl phosphatidylethanolamine and in anotherembodiment is dimyristoyl phosphatidylethanolamine.

In one embodiment, the dermatologic condition is a dermatologic disease.In another embodiment, the dermatologic condition is psoriasis. Inanother embodiment, the dermatologic condition is contact dermatitis. Inanother embodiment, the dermatologic condition is seborrheic dermatitis.In another embodiment, the dermatologic condition is atopic dermatitis.

BRIEF DESCRIPTION OF FIGURES

FIG. 1.1: Effect of Compound XXVI on the proliferation of cultured humanpsoriatic fibroblasts and Swiss 3T3 cells.

FIG. 1.2: Improvement of contact dermatitis score on vehicle- andCompound XXII-treated hands.

FIG. 2.1: A Lipid-conjugate protects BGM cells from membrane lysisinduced by combined action of hydrogen peroxide produced by glucoseoxidase (GO) and exogenous phospholipase A₂ (PLA₂).

FIG. 2.2: A Lipid-conjugate protects BGM cells from glycosaminoglycandegradation by hydrogen peroxide produced by glucose oxidase (GO).

FIG. 2.3: A Lipid-conjugate protects LDL from copper-induced oxidation.

FIG. 4.1A: Inhibition of the PLA₂ enzyme by Compound XXII.

FIG. 4.1B: Inhibition of the PLA₂ enzyme by Compound XXV.

DETAILED DESCRIPTION OF THE INVENTION

In another embodiment, the invention provides a method of treating adermatologic condition in a human subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof.

In another embodiment, the invention provides a method of treating adermatologic condition in a human subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof, wherein X is hyaluronic acid and L is        phosphatidylethanolamine.

In one embodiment, the invention provides a method of preventing adermatologic condition in a subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof.

In another embodiment, the invention provides a method of preventing ortreating contact dermatitis in a subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof.

In another embodiment, the invention provides a method of preventing ortreating atopic dermatitis in a subject, comprising the step ofadministering to said subject a compound represented by the structure ofthe general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000,        and/or a pharmaceutically acceptable salt or a pharmaceutical        product thereof.

In one embodiment, X in general formula (A) represents a polysaccharide.In one embodiment, the polysaccharide is carboxymethylcellulose, inanother embodiment, the polysaccharide is dextran, while in anotherembodiment, the polysaccharide is a glycosaminoglycan. In oneembodiment, the glycosaminoglycan is hyaluronic acid, in anotherembodiment, the glycosaminoglycan is chondroitin sulfate, while inanother embodiment, the glycosaminoglycan is heparin. In one embodiment,L in general formula (A) is phosphatidylethanolamine, which in oneembodiment is dipalmitoyl phosphatidylethanolamine and in anotherembodiment is dimyristoyl phosphatidylethanolamine.

In one embodiment, the methods of the present invention may be used toprevent or treat any dermatologic condition. In one embodiment, adermatological condition is a dermatologic disease, which in oneembodiment is dermatitis. In one embodiment, dermatitis is referred toas eczema. In another embodiment, the dermatologic condition is contactdermatitis, which in one embodiment is allergic contact dermatitis andin another embodiment is irritant contact dermatitis. In anotherembodiment, the dermatologic condition is atopic dermatitis, which inanother embodiment is infantile eczema, Besnier's prurigo, allergicdermatitis, flexural eczema, or disseminated neurodermatitis. In anotherembodiment, the dermatologic condition is seborrheic (or seborrhoeic)dermatitis, which in one embodiment is infantile seborrheic dermatitis,while in another embodiment, it's adult seborreic dermatitis. In anotherembodiment, the dermatologic condition is psoriasis. In anotherembodiment, a dermatologic condition is: neurodermatitis, scabies,systemic dermatitis, dermatitis herpetiformis, perioral dermatitis,discoid eczema, Nummular dermatitis, Housewives' eczema, Pompholyxdyshidrosis, Recalcitrant pustular eruptions of the palms and soles,Barber's or pustular psoriasis, Generalized Exfoliative Dermatitis,Stasis Dermatitis, varicose eczema, Dyshidrotic eczema, Lichen SimplexChronicus (Localized Scratch Dermatitis; Neurodermatitis), LichenPlanus, Fungal infection, which in one embodiment may be Candidaintertrigo, tinea capitis, white spot, panau, ringworm, athlete's foot,moniliasis, candidiasis; dermatophyte infection, vesicular dermatitis,chronic dermatitis, spongiotic dermatitis, dermatitis venata, Vidal'slichen, asteatosis eczema dermatitis, autosensitization eczema, or acombination thereof.

In another embodiment, the methods of the present invention may be usedto prevent or treat pimples, acne vulgaris, birthmarks, freckles,tattoos, scars, burns, sun burns, wrinkles, frown lines, crow's feet,café-au-lait spots, benign skin tumors, which in one embodiment, isSeborrhoeic keratosis, Dermatosis papulosa nigra, Skin Tags, Sebaceoushyperplasia, Syringomas, Xanthelasma, or a combination thereof; benignskin growths, viral warts, diaper candidiasis, folliculitis, furuncles,boils, carbuncles, fungal infections of the skin, guttate hypomelanosis,hair loss, impetigo, melasma, molluscum contagiosum, rosacea, scapies,shingles, erysipelas, erythrasma, herpes zoster, varicella-zoster virus,chicken pox, skin cancers, which in one embodiment, may be squamos cellcarcinoma, basal cell carcinoma, malignant melanoma; premalignantgrowths, which in one embodiment may be congenital moles, actinickeratosis; urticaria, hives, vitiligo, Ichthyosis, Acanthosis Nigricans,Bullous Pemphigoid, Corns and Calluses, Dandruff, Dry Skin, ErythemaNodosum, Graves' Dermopathy, Henoch-Schönlein Purpura, KeratosisPilaris, Lichen Nitidus, Lichen Planus, Lichen Sclerosus, Mastocytosis,Molluscum Contagiosum, Pityriasis Rosea, Pityriasis Rubra Pilaris,PLEVA, or Mucha-Habermann Disease, Epidermolysis Bullosa, SeborrheicKeratoses, Stevens-Johnson Syndrome, Pemphigus, or a combinationthereof.

In another embodiment, the methods of the present invention may be usedto prevent or treat insect bites or stings, which in one embodimentcomprise mosquito, tick, louse, flea, bed bug, which in one embodimentis Cimex lectularius or Cimex hemipterus in warmer areas, ant, snake,lizard (which in one embodiment are Gila monster Heloderma suspectum orthe beaded lizard H. horridum), spider, Black Widow Spider, BrownRecluse, Mite, Scorpion bites, or the like, or the skin reaction that issecondary to such bites. In one embodiment, stings comprise Bee, HoneyBee, Hornet, Wasp, or Yellow Jacket stings. In another embodiment, themethods of the present invention may be used to prevent or treat bitesor stings from marine animals, including Coelenterates, including thecorals, sea anemones, jellyfishes, and hydroids (eg, Portugueseman-of-war); stingrays; mollusks, including the cones, which is oneembodiment is Conus californicus, octopuses, and bivalves; andEchinoderms and sea urchins which in one embodiment are Globiferouspedicellariae.

In another embodiment, the methods of the present invention may be usedto prevent or treat dermatologic conditions that are associated with theeye area, which in one embodiment may be Syringoma, Xanthelasma,Impetigo, atopic dermatitis, contact dermatitis, or a combinationthereof; the scalp, fingernails, which in one embodiment may beinfection by bacteria, fungi, yeast and virus, Paronychia, or psoriasis;mouth area, which in one embodiment is Oral Lichen Planus, Cold Sores(Herpetic Gingivostomatitis), Oral Leukoplakia, Oral Candidiasis, or acombination thereof; or a combination thereof. In one embodiment, themethods of the present invention may be used to prevent or treatdermatologic conditions in babies, infants, toddlers, young children,older children, adolescents, young adults, adults, elderly, or acombination thereof.

It is to be understood that the methods of the present invention may beused to prevent or treat any inflammatory skin condition.

In one embodiment, “preventing, or treating” refers to any one or moreof the following: delaying the onset of symptoms, reducing the severityof symptoms, reducing the severity of an acute episode, reducing thenumber of symptoms, reducing the incidence of disease-related symptoms,reducing the latency of symptoms, ameliorating symptoms, reducingsecondary symptoms, reducing secondary infections, prolonging patientsurvival, preventing relapse to a disease, decreasing the number orfrequency of relapse episodes, increasing latency between symptomaticepisodes, increasing time to sustained progression, expeditingremission, inducing remission, augmenting remission, speeding recovery,or increasing efficacy of or decreasing resistance to alternativetherapeutics. In one embodiment, “treating” refers to both therapeutictreatment and prophylactic or preventative measures, wherein the objectis to prevent or lessen the targeted pathologic condition or disorder asdescribed hereinabove.

In one embodiment, the compounds for use in the methods of the presentinvention may be used to address the symptoms of a dermatologiccondition, which in one embodiment are primary, while in anotherembodiment, are secondary to the dermatologic condition. In oneembodiment, “primary” refers to a symptom that is a direct result of thedermatologic condition, while in one embodiment, “secondary” refers to asymptom that is derived from or consequent to a primary cause.

In another embodiment, “symptoms” may be any manifestation of a diseaseor pathological condition, comprising inflammation, swelling,superficial skin inflammation, presence of vesicles, poorly marginatedredness, red skin, dry skin, thick skin, leathery skin, edema, oozing,crusting, scaling, burning, stinging, pruritus, itching,lichenification, blisters, papules, vesicles, bullae, blisters,tenderness, skin lesion, which in one embodiment, may be oozing,draining, crusting, scaly, raw, thickened or a combination thereof,rash, fever, pain, bleeding or a combination thereof.

As described hereinabove, in one embodiment, the methods of the presentinvention prevent or treat contact dermatitis. In one embodiment,contact dermatitis is an inflammation produced by substances contactingthe skin and causing toxic (irritant) or allergic reactions. In oneembodiment, contact dermatitis is initiated by plants (in oneembodiment, poison ivy, poison oak, etc.), sensitizers used in themanufacture of shoes and clothing, metal compounds, dyes, cosmetics,industrial agents, rubber accelerators, latex gloves, latex condoms,ingredients in topical drugs. In one embodiment, contact dermatitis isphotoallergic or phototoxic contact dermatitis in which topicallyapplied dermatitides evoke a response after being exposed to light. Inone embodiment, aftershave lotions, sunscreens, and topical sulfonamidesare commonly responsible for photoallergic contact dermatitis. Inanother embodiment, perfumes, coal tar, psoralens, and oils used inmanufacturing are commonly responsible for phototoxic contactdermatitis. In one embodiment, symptoms of contact dermatitis comprisetransient redness, swelling, bullae, erythema, blisters, vesicles whichmay rupture, ooze, or crust, scaling, skin thickening, acuteinflammation, chronic inflammation, or a combination thereof.

As described hereinabove, in one embodiment, the methods of the presentinvention prevent or treat atopic dermatitis, which in one embodiment,is chronic, pruritic, superficial inflammation of the skin, frequentlyassociated with a personal or family history of allergic disorders(e.g., hay fever, asthma). In one embodiment, susceptibility to atopicdermatitis is genetic, is triggered by environmental agents and factors,is auto-immune, or a combination thereof. In one embodiment, sufferersof atopic dermatitis have high serum levels of reaginic (IgE)antibodies, peripheral eosinophilia, high levels of cAMPphosphodiesterase in their white blood cells, or a combination thereof.In one embodiment, symptoms of atopic dermatitis in infants comprisered, weeping, and/or crusted lesions on the face, scalp, diaper area andextremities. In another embodiment, symptoms comprise erythema,lichenification in the antecubital and popliteal fossae and on theeyelids neck, or wrists, pruritus, atopic pleat (Dennie-Morgan fold),hyperlinear palms, hyperpigmented eyelids, ichthyosis, keratosispilaris, urticaria, or a combination thereof. In one embodiment, atopicdermatitis appears with and/or may be influenced by secondary bacterialinfections, regional lymphadenitis, contact dermatitis from topicalallergens, emotional stress, ambient temperature and/or humiditychanges, fragrances, fabric softeners, wool garments, house mites, or acombination thereof.

As described hereinabove, in one embodiment, the methods of the presentinvention prevent or treat seborrheic dermatitis, which in oneembodiment is an inflammatory scaling disease or the scalp, face, orother areas with high densities or large oil glands. In one embodiment,symptoms of seborrheic dermatitis comprise dry or greasy diffuse scalingof the scalp (dandruff) with variable pruritus, yellow-red scalingpapules along the hairline, behind the ears, in the external auditorycanals, on the eyebrows, on the bridge of the nose, in the nasolabialfolds over the sternum, or a combination thereof, marginal blepharitiswith dry yellow crusts, conjunctival irritation, or a combinationthereof. In one embodiment, seborrheic dermatitis in newborns may becalled cradle cap and, in one embodiment comprises symptoms of thick,yellow, crusted scalp lesion, fissuring, yellow scaling behind the ears,red facial papules, stubborn diaper rash, or a combination thereof. Inone embodiment, genetic factors, emotional or physical stress, climate,season, or a combination thereof may affect onset or progression ofsymptoms. In one embodiment, seborrheic dermatitis may precede or beassociated with the development of psoriasis. In one embodiment, themethods of the present invention prevent or treat seborrheic dermatitisin patients with neurologic disease (in one embodiment, Parkinson'sDisease) or Human Immunodeficiency Virus (HIV).

As described hereinabove, in one embodiment, the methods of the presentinvention prevent or treat psoriasis, which, in one embodiment, is anoncontagious inflammatory skin disease characterized by recurringreddish patches covered with silvery scales. In one embodiment, thethick scaling is attributed to increased epidermal cell proliferationand concomitant dermal inflammation. In some embodiments, the methods ofthe present invention may treat psoriatic arthritis, erythrodermicpsoriasis, or pustular psoriasis, which in one embodiment may be vonZumbusch type psoriasis, Palmo-plantar pustulosis, Acropustulosis(acrodermatitis continua of Hallopeau), or Barber's psoriasis. In oneembodiment, the methods of the present invention prevent or treat plaquepsoriasis, guttate psoriasis, inverse psoriasis, erythrodermicpsoriasis, a combination thereof, or other forms of psoriasis that areknown in the art. In one embodiment, symptoms of psoriasis comprisesharply demarcated lesions, variously pruritic lesions, ovoid orcircinate erythematous papules, plaques covered with overlapping thicksilvery micaceous or slightly opalescent shiny scales, papules thatextend and coalesce to produce large plaques in annular and gyratepatterns, stippling, pitting, fraying, discoloration or separation ofthe distal and/or lateral margins of the nail plate (onycholysis),thickening, with hyperkeratotic debris under the nail plate, or acombination thereof. In one embodiment, psoriasis has chronic remissionsand recurrences that vary in frequency and duration. In one embodiment,factors that precipitate the onset or severity of psoriasis includelocal trauma, irritation, severe sunburn, viremia, allergic drugreactions, topical and systemic drugs (which in one embodiment comprisechloroquine antimalarial therapy, lithium, β-blockers, interferon-α, ora combination thereof), withdrawal of systemic corticosteroids, or acombination thereof. In one embodiment, psoriasis is found on the scalp(in one embodiment, the postauricular regions), extensor surface ofextremities (in one embodiment, elbows, knees, or a combinationthereof), face, nails, sacral area, buttocks, genitals, or combinationthereof.

In one embodiment, the methods of the present invention may be used totreat a dermatologic condition in a subject that is immunosuppressed,while in another embodiment, in a subject that is immunodeficient.

In one embodiment, the methods of the present invention may be used totreat a dermatologic condition in a subject of any species, which in oneembodiment is a mammal, while in another embodiment, the subject isnon-mammalian. In one embodiment, the subject is a vertebrate. In oneembodiment, the subject is a human. In other embodiments the subject isa primate, equine, avian, bovine, ovine, caprine, porcine, canine,feline or murine subject.

In one embodiment, the methods of the present invention may be used totreat a veterinary dermatologic condition, which in one embodiment iseczematoid dermatitis, chronic dermatitis, equine exuberant granuloma(“proud flesh”), decubitis ulcers, canine cutaneous granulomas (“lick”granuloma), and others that are known to one of ordinary skill in theart.

In one embodiment, the methods of the present invention may be used totreat a dermatologic condition in a subject anywhere on the body of thesubject, including, inter alia, scalp, face, head, neck, shoulders,arms, elbows, behind the elbows, wrists, hands, fingers, palms, trunk,back, chest, shoulders, buttocks, genitals, legs, knees, behind theknees, feet, toes, ankles, soles, etc.

In one embodiment, the compounds for use in the present invention (fore.g., a lipid or phospholipid moiety bound to a physiologicallyacceptable monomer, dimer, oligomer, or polymer) are referred to hereinas “Lipid-conjugates”.

In one embodiment, Lipid-conjugates of the present invention prevent ortreat a dermatologic condition. In another embodiment, Lipid-conjugatesprevent and treat a skin hypersensitivity reaction. This is exemplifiedin Tables 1.1-1.4 and represents an embodiment of this invention. Inanother embodiment, Lipid-conjugates are effective in treatingpsoriasis, as exemplified in FIG. 1.2. In another embodiment,Lipid-conjugates are effective in treating contact dermatitis in humans,as exemplified in FIG. 1.2. In another embodiment, Lipid-conjugatesdecrease ear swelling in an animal model of skin hypersensitivityreaction, when administered i.p. (Table 1.1), s.c. (Table 1.2, andtopically bilaterally (Table 1.3) or unilaterally (Table 1.4). Inanother embodiment, Lipid-conjugates inhibit proliferation of psoriaticskin fibroblasts and Swiss 3T3 cells (as exemplified in FIG. 1.1). Inanother embodiment, Lipid-conjugates decrease contact dermatitis scores(as exemplified in FIG. 1.2). In one embodiment, Compound XXVI (seecompound descriptions hereinbelow) is useful to treat dermatologicconditions. In another embodiment, Compound XXVI decreases swellingafter i.p. or s.c. administration, as exemplified in Tables 1.1 and 1.2,and inhibits the proliferation of cultured psoriatic skin fibroblastsand Swiss 3T3 cells, as exemplified in FIG. 1.1. In another embodiment,Compound XXIX (see compound descriptions hereinbelow) is useful to treatdermatologic conditions. In another embodiment, Compound XXIX decreasesswelling after bilateral or unilateral topical administration, asexemplified in Tables 1.3 and 1.4. In another embodiment, Compound XXII(see compound descriptions hereinbelow) is useful to treat dermatologicconditions. In another embodiment, Compound XXII decreases contactdermatitis scores in patients after unilateral topical administration,as exemplified in FIG. 1.2.

In one embodiment, the invention provides a method for treating asubject with a dermatologic condition marked by unchecked inflammation,inappropriate cytokine response, or a combination thereof. In anotherembodiment, local cytokine profiles can be altered by the treatmentaccording to the methods of the present invention. In one embodiment,the determination of the modulation of cytokines may be performed asdescribed by U.S. application Ser. No. 10/952,496 filed Sep. 29, 2004,which is incorporated herein by reference in its entirety.

In another embodiment, the compounds for use in the present inventionmay be used to treat conditions secondary to dermatologic conditions,including sepsis and oxidative injury. The use of Lipid-conjugates totreat sepsis is demonstrated in U.S. application Ser. No. 10/627,981,filed Jul. 28, 2003, which is incorporated herein by reference in itsentirety.

Administration of the Lipid-conjugates in a diversity of animal and cellmodels of disease invoked remarkable, and unexpected, cytoprotectiveeffects, which, as exemplified herein, are useful in the prevention andtreatment of dermatologic conditions and related diseases and/orconditions. Lipid-conjugates reduce ear swelling in mice induced byoxazolone, inhibit proliferation of cultured psoriatic skin fibroblastsand Swiss 3T3 cells, and reduce symptoms of contact dermatitis in humans(as exemplified in Example 1). In one embodiment, Lipid-conjugates mayalso prevent and/or treat dermatologic conditions via protecting againstoxidative injury (as exemplified in Example 2) and via theirmembrane-stabilizing effects (as exemplified in Example 3). Finally, inone embodiment, Lipid-conjugates may prevent or treat dermatologicconditions via their PLA₂ enzyme inhibitory activity (as exemplified inExample 4).

The compounds for the use in the present invention also reduce sPLA2expression in rat lung, reduce cysteinyl leukotrienes, reduce NOproduction, prevent airway remodeling, and reduce tumor necrosisfactor-α (TNF-α) in animal and cell models of obstructive respiratorydisease; inhibit NO production, PGE2, sPLA2, and oleic acid release fromPC12 and glial cells in cell models of CNS injury; reduce IL-8, Gro-α,ena-78, and NF-κB in an in vitro model of acute respiratory distresssyndrome (as exemplified in U.S. application Ser. No. 10/952,496, whichis incorporated by reference herein).

In one embodiment of the present invention, the useful pharmacologicalproperties of the Lipid-conjugates, some of which are describedhereinabove, may be applied for clinical use, and disclosed herein asmethods for the prevention or treatment of a disease. The biologicalbasis of these methods may be readily demonstrated by standard cellularand animal models of disease, for example, as described in the Examples1-5, hereinbelow.

In one embodiment, the pharmacological activities of Lipid-conjugates,including membrane stabilization, anti-inflammation, anti-oxidantaction, and attenuation of chemokine levels, may contribute to aLipid-conjugate-treated cell's resistance to dermatologic conditions ordiseases. In one embodiment, cell membrane stabilization may ameliorateor prevent tissue injury arising in the course of a dermatologiccondition. In another embodiment, anti-oxidant action may limitoxidative damage to cell and blood components arising in the course of adermatologic condition. In another embodiment, attenuation of chemokineslevels may attenuate physiological reactions to stress that arise in thecourse of a dermatologic condition.

The combination of lipids, such as, but not limited tophosphatidylethanolamine and phosphatidylserine, with additional monomeror polymer moieties, is thus a practical route to the production of newdrugs for medical purposes, provided that the resultant chemicalcomposition displays the desired range of pharmacological properties. Inone embodiment, the Lipid-conjugates of this invention possess acombination of multiple and potent pharmacological effects in additionto the ability to inhibit the extracellular form of the enzymephospholipase A2. While the pharmacological activity of theLipid-conjugates described herein may be due in part to the nature ofthe lipid moiety, the multiple and diverse combination ofpharmacological properties observed for the Lipid-conjugates emergesfrom the ability of the compound structure to act essentially as severaldifferent drugs in one chemical entity.

In the cases described herein, the diversity of biological activitiesand the effectiveness in disease exhibited by the compounds for use inthe present invention far exceed the properties anticipated by use ofthe starting materials themselves, when administered alone or incombination. However, the phospholipid conjugate compounds, alone or incombination, are valuable when used in the methods of treating diseasesand conditions specifically described herein.

In one embodiment, methods of the present invention involve treating asubject by inter alia controlling the expression, production, andactivity of phospholipases such as PLA2; controlling the productionand/or action of lipid mediators, such as eicosanoids, plateletactivating factor (PAF) and lyso-phospholipids; amelioration of damageto cell surface glycosaminoglycans (GAG) and proteoglycans; controllingthe production of oxidants, oxygen radicals and nitric oxide; protectionof cells, tissues, and plasma lipoproteins from damaging agents, such asreactive oxygen species (ROS) and phospholipases; controlling theexpression, production, and activity of cytokines, chemokines andinterleukins; anti-oxidant therapy; anti-endotoxin therapy or anycombination thereof.

In one embodiment of the invention, the term “controlling” refers toinhibiting the production and action of the above mentioned factors inorder to maintain their activity at the normal basal level and suppresstheir activation in pathological conditions.

In one embodiment of the invention, a dermatologic condition ischaracterized by the presence of damaging agents, which comprise, interalia, phospholipases, reactive oxygen species (ROS), free radicals,lysophospholipids, fatty acids or derivatives thereof, hydrogenperoxides, phospholipids, oxidants, cationic proteins, streptolysins,proteases, hemolysins, or sialidases.

Dosages and Routes of Administration

This invention encompasses administration of compounds as describedherein or compositions comprising the same, for treating psoriasis,dermatitis, or other dermatologic conditions.

In one embodiment, compositions of this invention are pharmaceuticallyacceptable. In one embodiment, the term “pharmaceutically acceptable”refers to any formulation which is safe, and provides the appropriatedelivery for the desired route of administration of an effective amountof at least one compound for use in the present invention. This termrefers to the use of buffered formulations as well, wherein the pH ismaintained at a particular desired value, ranging from pH 4.0 to pH 9.0,in accordance with the stability of the compounds and route ofadministration.

In one embodiment, a Lipid-conjugate used in the methods of thisinvention may be administered alone or within a composition. In anotherembodiment, compositions comprising Lipid-conjugates in admixture withconventional excipients, i.e. pharmaceutically acceptable organic orinorganic carrier substances suitable for parenteral, enteral (e.g.,oral) or topical application which do not deleteriously react with theactive compounds may be used. In one embodiment, suitablepharmaceutically acceptable carriers include but are not limited towater, salt solutions, alcohols, gum arabic, vegetable oils, benzylalcohols, polyethylene glycols, gelatine, carbohydrates such as lactose,amylose or starch, magnesium stearate, talc, silicic acid, viscousparaffin, white paraffin, glycerol, alginates, hyaluronic acid,collagen, perfume oil, fatty acid monoglycerides and diglycerides,pentaerythritol fatty acid esters, hydroxy methylcellulose, polyvinylpyrrolidone, etc. In another embodiment, the pharmaceutical preparationscan be sterilized and if desired mixed with auxiliary agents, e.g.,lubricants, preservatives, stabilizers, wetting agents, emulsifiers,salts for influencing osmotic pressure, buffers, coloring, flavoringand/or aromatic substances and the like which do not deleteriously reactwith the active compounds. In another embodiment, they can also becombined where desired with other active agents, e.g., vitamins.

In one embodiment, the therapeutic compositions of the present inventioncomprise a Lipid-conjugate and one or more additional compoundseffective in preventing or treating dermatologic conditions. In oneembodiment, the additional compound is a moisturizer or an emollient,which in one embodiment is petrolatum, white petrolatum, hydrogenatedvegetable oil, hydrophilic petrolatum, panthenol, primrose oil, omega-3fish oils, omega-6 fish oils, linoleic acid, flax seed oil, ceramide,borage oil (linoleic acid), tocopherol (Vitamin E), tocopherollinoleate, dimethicone, glycerine or a combination thereof. In oneembodiment, moisturizers improve the ability of the skin to absorb otheradministered compounds, including inter alia, the compounds for use inthe present invention. In another embodiment, moisturizing agentsminimize or prevent the skin from drying and cracking, therebydecreasing susceptibility of skin to environmental factors that generatefree radicals, thereby preventing additional damage to the skin.

In another embodiment, the additional compound is a topical steroid,which in one embodiment is hydrocortisone, in one embodiment 1%hydrocortisone, triamcinolone, fluocinolone acetonide, halcinonide,halobetasol propionate, clobetasol propionate, betamethasonedipropionate, betamethasone valerate, and triamcinolone acetonide or acombination thereof; oral steroids; topical immunomodulators including,inter alia, tacrolimus, pimecrolimus, Ascomycin, cyclosporine, or acombination thereof; antihistamines, which in one embodiment ishydroxyzine or diphenhydramine hydrochloride, Ketotifen, Doxepin;biologics, which in one embodiment comprises Amevive (alefacept),Enbrel, Humira, Raptiva, Remicade, or a combination thereof; or acombination thereof. In another embodiment, the additional compound isan antibiotic, which in one embodiment comprise cloxacillin, cephalexin,penicillin, clindamycin or a combination thereof. In another embodiment,the additional compound is methotrexate, tar, coal tar, anthralin,dovonex, salicyclic acid, tazorac, moisturizers, aloe vera, soriatane,accutane, hydrea, mycophenolate mofetil, sulfasalazine, 6-thioguanine,or a combination thereof. In another embodiment, additional compoundscomprise acyclovir, which in one embodiment is particularly effective inpatients with eczema herpeticum. In one embodiment, additional compoundsto treat seborrheic dermatitis comprise zinc pyrithione, seleniumsulfide, sulfur, tar shampoo, flucinolone acetonide solution,triamcinolone acetonide lotion, ketoconazole cream, other imidazoles, ora combination thereof.

In another embodiment, the additional compound is an anti-inflammatoryagent, which in one embodiment comprises aspirin, ibuprofen, ketoprofen,naproxen, or a combination thereof.

In another embodiment, the additional compound is an exfoliant, which inone embodiment comprises an enzymatic exfoliant or a mono- or-poly-hydroxy acid. In one embodiment, the exfoliant is an alpha-hydroxyacid, beta-hydroxy acid, tannic acid, glycolic acid, lactic acid, citricacid, salicylic acid, or a combination thereof. In another embodiment,the additional compound is an analgesic, or anesthetic, while in anotherembodiment it is aloe vera gel, aloe vera, licorice extract, pilewort,Canadian willow root, zinc, allantoin, or a combination thereof. Inanother embodiment, the additional compound is an anti-oxidant.

In another embodiment, the additional compound is an antibacterial,antifungal, antiviral, antihelminthic agent, or a combination thereof.In one embodiment, the additional compound is echinacea, golden seal,benzalkonium chloride, benzethonium chloride, iodine, grape seedextract, pomegranate extract, green tea extract or polyphenols, and thelike, or combinations thereof. In one embodiment, an antihelminthicagent is metronidazole. In one embodiment, antiviral agent is acyclovir,tamvir, penciclovir, or a combination thereof. In one embodiment, theantibacterial agent is triclosan, neomycin, polymyxin, bacitracin,clindamycin, benzoyl peroxide, a tetracycline, a sulfa drug, apenicillin, a quinolone, a cephalosporin, or a combination thereof. Inone embodiment, the antifungal agent is famesol, econazole, fluconazole,clotrimazole, ketoconazole, calcium or zinc undecylenate, undecylenicacid, butenafine hydrochloride, ciclopirox olaimine, miconazole nitrate,nystatin, sulconazole, terbinafine hydrochloride, and the like, or acombination thereof.

In one embodiment, the therapeutic compositions of the present inventionare administered with other treatments that relieve symptoms. In oneembodiment, other treatments comprise wet wrap bandaging, phototherapy,UV light exposure (UVA, UVB, or a combination thereof), psoralen plusUV-A (PUVA), UV-B 1 (narrow band UV-B) therapy, Balneotherapy,Climatotherapy, aromatherapy, relaxation, homeopathy, chinese herbaltreatments, oil of evening primrose, and the like.

In one embodiment, the route of administration may be parenteral,enteral, or a combination thereof. In another embodiment, the route maybe intra-ocular, conjunctival, topical, transdermal, intradermal,subcutaneous, intraperitoneal, intravenous, intra-arterial, vaginal,rectal, intratumoral, parcanceral, transmucosal, intramuscular,intravascular, intraventricular, intracranial, inhalation, nasalaspiration (spray), sublingual, oral, aerosol or suppository or acombination thereof. In one embodiment, the dosage regimen will bedetermined by skilled clinicians, based on factors such as exact natureof the condition being treated, the severity of the condition, the ageand general physical condition of the patient, body weight, and responseof the individual patient, etc.

For topical application, particularly in the area around the eye, anadmixture of the compounds with conventional creams, lotions, or delayedrelease patches is acceptable. Such a cream or lotion may comprise anyagent described herein, and, in one embodiment, may be used to treatskin diseases and/or dermatologic conditions, such as, in oneembodiment, contact dermatitis, atopic dermatitis, psoriasis, or acombination thereof.

Suitable dosage forms for topical administration include, but are notlimited to, dispersions, lotions; creams; gels; pastes; powders; aerosolsprays; syrups or ointments on sponges or cotton applicators; andsolutions or suspensions in an aqueous liquid, non-aqueous liquid,oil-in-water emulsion, or water-in-oil liquid emulsion. Because of itsease of administration, a cream, lotion, or ointment represents the mostadvantageous topical dosage unit form, in which case liquidpharmaceutical carriers may be employed in the composition. Thesecreams, lotions, or ointments, may be prepared as rinse-off or leave-onproducts, as well as two stage treatment products for use with otherskin cleansing or managing compositions. In a preferred embodiment, thecompositions are administered as a rinse-off product in a higherconcentration form, such as a gel, and then a leave-on product in alower concentration to avoid irritation of the skin. Each of these formsis well understood by those of ordinary skill in the art, such thatdosages may be easily prepared to incorporate the pharmaceuticalcomposition of the invention.

Ointment preparations may be roughly classified into fat/oil typeointments, emulsified ointments, water-soluble ointments and suspendedointments according to the type of the base (vehicle) used therefor. Anointment may comprise, for example, fats, fatty oils, lanolin, vaseline,paraffins, waxes, resins, plastics, glycols, higher alcohols, glycerol,water, emulsifiers, suspending agents or other appropriate additives asa diluent, carrier or as a vehicle. Manufacture of an ointmentcomprises, for example, adding the compound of the present invention tothe appropriate additives, diluents, carriers or vehicles followed bymixing to make the mixture homogeneous.

For parenteral application, particularly suitable are injectable,sterile solutions, preferably oily or aqueous solutions, as well assuspensions, emulsions, or implants, including suppositories and enemas.Ampoules are convenient unit dosages. Such a suppository may compriseany agent described herein.

For application by inhalation, solutions or suspensions of the compoundsmixed and aerosolized or nebulized in the presence of the appropriatecarrier suitable. Such an aerosol may comprise any agent describedherein.

For enteral application, particularly suitable are tablets, dragees,liquids, drops, or capsules. In one embodiment, a sweetened vehicle isemployed when a syrup, elixir, or the like is used for enteralapplication.

Sustained or directed release compositions can be formulated, e.g.,liposomes or those wherein the active compound is protected withdifferentially degradable coatings, e.g., by microencapsulation,multiple coatings, etc. It is also possible to freeze-dry the newcompounds and use the lyophilisates obtained, for example, for thepreparation of products for injection.

Thus, in one embodiment, the route of administration may be directed toan organ or system that is affected by dermatologic conditions. Forexample, compounds may be administered topically to treat dermatologicconditions. In another embodiment, the route of administration may bedirected to a different organ or system than the one that is affected bydermatologic conditions. For example, compounds may be administeredparenterally to treat dermatologic conditions. Thus, the presentinvention provides for the use of Lipid-conjugates in various dosageforms suitable for administration using any of the routes listedhereinabove.

In general, the doses utilized for the above described purposes willvary, but will be in an effective amount to exert the desired effect. Asused herein, the term “pharmaceutically effective amount” refers to anamount of a compound of formulae A and I-LXXXVII as describedhereinbelow, which will produce the desired alleviation in symptoms orother desired phenotype in a patient. The doses utilized for any of theabove-described purposes will generally be from 1 to about 1000milligrams per kilogram of body weight (mg/kg), administered one to fourtimes per day, or by continuous IV infusion. In one embodiment, atopical daily dose range, in single or divided doses, for the conditionsdescribed herein is from about 1 mg to 20,000 mg, more preferably about2,000 mg to 16,000 mg, and most preferably about 6,000 mg to 10,000 mgof the active components (i.e., excluding excipients and carriers). Whenthe compositions are dosed topically or intraocularly, they willgenerally be in a concentration range of from 0.1 to about 10% w/v,administered 1-4 times per day. In one embodiment, the compositions foruse in the methods of the present invention are administered topicallytwo times a day in a concentration of 1% and the following w/w % ratios:Water 70.0, Cetyl Alcohol, 10.6, Paraffin, White soft 10.6, PropyleneGlycol 7.2, HyPE 1.0 and Sodium Dodecyl Sulfate 0.6 (as exemplified inExample 1.6).

In one embodiment of the invention, the concentrations of the compoundswill depend on various factors, including the nature of the condition tobe treated, the condition of the patient, the route of administrationand the individual tolerability of the compositions.

It will be appreciated that the actual preferred amounts of activecompound in a specific case will vary according to the specific compoundbeing utilized, the particular compositions formulated, the mode ofapplication, and the particular conditions and organism being treated.Dosages for a given host can be determined using conventionalconsiderations, e.g., by customary comparison of the differentialactivities of the subject compounds and of a known agent, e.g., by meansof an appropriate, conventional pharmacological protocol.

In one embodiment, the compounds of the invention may be administeredacutely for acute treatment of temporary conditions, or may beadministered chronically, especially in the case of progressive,recurrent, or degenerative disease. In one embodiment, one or morecompounds of the invention may be administered simultaneously, or inanother embodiment, they may be administered in a staggered fashion. Inone embodiment, the staggered fashion may be dictated by the stage orphase of the disease.

In one embodiment, the present invention offers methods for thetreatment of disease based upon administration of lipids covalentlyconjugated through their polar head group to a physiologicallyacceptable chemical moiety, which may be of high or low molecularweight.

The present invention has been illustrated in terms of the anti-diseaseactivity of Lipid-conjugates and methods of their use as pharmaceuticalcompositions in the treatment of disease. The following sections presentsome examples of the therapeutic Lipid-conjugate compounds for use inthe present invention and their chemical preparation.

Compounds

In one embodiment, the compounds for use in the present inventioncomprise a lipid or phospholipid moiety bound to a physiologicallyacceptable monomer, dimer, oligomer, or polymer. In one embodiment, thelipid compounds (Lipid-conjugates) for use in the present invention aredescribed by the general formula:

[phosphatidylethanolamine-Y]n-X

[phosphatidylserine-Y]n-X

[phosphatidylcholine-Y]n-X

[phosphatidylinositol-Y]n-X

[phosphatidylglycerol-Y]n-X

[phosphatidic acid-Y]n-X

[lyso-phospholipid-Y]n-X

[diacyl-glycerol-Y]n-X

[monoacyl-glycerol-Y]n-X

[sphingomyelin-Y]n-X

[sphingosine-Y]n-X

[ceramide-Y]n-X

wherein

-   Y is either nothing or a spacer group ranging in length from 2 to 30    atoms; and-   X is a physiologically acceptable monomer, dimer, oligomer or    polymer; and-   n is the number of lipid molecules bound to a molecule of X, wherein    n is a number from 1 to 1000.

In one embodiment, the invention provides low-molecular weightLipid-conjugates, previously undisclosed and unknown to possesspharmacological activity, of the general formula described hereinabove.In another embodiment, wherein the general formula described hereinabovedescribes low-molecular weight Lipid-conjugates, X is a mono- ordisaccharide, carboxylated disaccharide, mono- or dicarboxylic acids, asalicylate, salicylic acid, aspirin, lactobionic acid, maltose, an aminoacid, glycine, acetic acid, butyric acid, dicarboxylic acid, glutaricacid, succinic acid, fatty acid, dodecanoic acid, didodecanoic acid,bile acid, cholic acid, cholesterylhemmisuccinate, a di- or tripeptide,an oligopeptide, a trisacharide, or a di- or trisaccharide monomer unitof heparin, heparan sulfate, keratin, keratan sulfate, chondroitin,chondroitin-6-sulfate, chondroitin-4-sulfate, dermatin, dermatansulfate, dextran, or hyaluronic acid.

In one embodiment of this invention, X is salicylate, salicylic acid,aspirin, a monosaccharide, lactobionic acid, maltose, an amino acid,glycine, carboxylic acid, acetic acid, butyric acid, dicarboxylic acid,glutaric acid, succinic acid, fatty acid, dodecanoic acid, didodecanoicacid, bile acid, cholic acid, cholesterylhemmisuccinate, a dipeptide, adisaccharide, a trisaccharide, an oligosaccharide, a polysaccharide, ahetero-polysaccharide, a homo-polysaccharide, a polypyranose, anoligopeptide, or a di- or trisaccharide monomer unit of heparin, heparansulfate, keratin, keratan sulfate, chondroitin, chondroitin-6-sulfate,chondroitin-4-sulfate, dermatin, dermatan sulfate, dextran, orhyaluronic acid, a glycosaminoglycan, polygeline (‘haemaccel’),alginate, hydroxyethyl starch (hetastarch), polyethylene glycol,polycarboxylated polyethylene glycol, chondroitin-6-sulfate,chondroitin-4-sulfate, keratin, keratin sulfate, heparan sulfate,dermatin, dermatan sulfate, carboxymethylcellulose, heparin, dextran, orhyaluronic acid.

In one embodiment, X is conjugated to the lipid, phospholipid, or spacervia an ester bond. In another embodiment, X is conjugated to the lipid,phospholipid, or spacer via an amide bond.

As defined by the structural formulae provided herein for theLipid-conjugates, these compounds may contain one to one thousand lipidmoieties bound to a single physiologically acceptable polymer molecule.In one embodiment of this invention, n is a number from 1 to 1000. Inanother embodiment, n is a number from 1 to 500. In another embodiment,n is a number from 1 to 100. In another embodiment, n is a number from 2to 1000. In another embodiment, n is a number from 2 to 100. In anotherembodiment, n is a number from 2 to 200. In another embodiment, n is anumber from 3 to 300. In another embodiment, n is a number from 10 to400. In another embodiment, n is a number from 50 to 500. In anotherembodiment, n is a number from 100 to 300. In another embodiment, n is anumber from 300 to 500. In another embodiment, n is a number from 500 to800. In another embodiment, n is a number from 500 to 1000.

In one embodiment of the invention, when the conjugated moiety is apolymer, the ratio of lipid moieties covalently bound may range from oneto one thousand lipid residues per polymer molecule, depending upon thenature of the polymer and the reaction conditions employed. For example,the relative quantities of the starting materials, or the extent of thereaction time, may be modified in order to obtain Lipid-conjugateproducts with either high or low ratios of lipid residues per polymer,as desired.

In one embodiment, the set of compounds comprisingphosphatidylethanolamine covalently bound to a physiologicallyacceptable monomer, dimer, oligomer, or polymer, is referred to hereinas the PE-conjugates. In one embodiment, the phosphatidylethanolaminemoiety is dipalmitoyl phosphatidylethanolamine. In another embodiment,the phosphatidylethanolamine moiety is dimyristoylphosphatidylethanolamine. In another embodiment, related derivatives, inwhich either phosphatidylserine, phosphatidylcholine,phosphatidylinositol, phosphatidic acid or phosphatidylglycerol areemployed in lieu of phosphatidylethanolamine as the lipid moiety provideequivalent therapeutic results, based upon the biological experimentsdescribed below for the Lipid-conjugates and the structural similaritiesshared by these compounds.

In another embodiment, the lipid or phospholipid moiety is phosphatidicacid, an acyl glycerol, monoacylglycerol, diacylglycerol,triacylglycerol, sphingosine, sphingomyelin, chondroitin-4-sulfate,chondroitin-6-sulfate, ceramide, phosphatidylethanolamine,phosphatidylserine, phosphatidylcholine, phosphatidylinositol, orphosphatidylglycerol, or an ether or alkyl phospholipid derivativethereof.

In one embodiment, Lipid-conjugate derivatives relevant to thisinvention are Lipid-conjugates wherein at least one of the fatty acidgroups of the lipid moieties at position C1 or C2 of the glycerolbackbone are substituted by a long chain alkyl group attached by amide,ether or alkyl bonds, rather than ester linkages.

In the methods, according to embodiments of the invention, theLipid-conjugates administered to the subject are comprised from at leastone lipid moiety covalently bound through an atom of the polar headgroup to a monomeric or polymeric moiety (referred to herein as theconjugated moiety) of either low or high molecular weight. When desired,an optional bridging moiety can be used to link the Lipid-conjugatesmoiety to the monomer or polymeric moiety. The conjugated moiety may bea low molecular weight carboxylic acid, dicarboxylic acid, fatty acid,dicarboxylic fatty acid, acetyl salicylic acid, cholic acid,cholesterylhemisuccinate, or mono- or di-saccharide, an amino acid ordipeptide, an oligopeptide, a glycoprotein mixture, a di- ortrisaccharide monomer unit of a glycosaminoglycan such as a repeatingunit of heparin, heparan sulfate, hyaluronic acid, chondroitin-sulfate,dermatan, keratan sulfate, or a higher molecular weight peptide oroligopeptide, a polysaccharide, a hetero-polysaccharide, ahomo-polysaccharide, a polypyranose, polyglycan, protein,glycosaminoglycan, or a glycoprotein mixture. The composition of somephospholipid-conjugates of high molecular weight, and associatedanalogues, are the subject of U.S. Pat. No. 5,064,817, which isincorporated herein in its entirety by reference.

In one embodiment, the term “moiety” means a chemical entity otherwisecorresponding to a chemical compound, which has a valence satisfied by acovalent bond.

In one embodiment, examples of polymers which can be employed as theconjugated moiety for producing Lipid-conjugates for use in the methodsof this invention may be physiologically acceptable polymers, includingwater-dispersible or -soluble polymers of various molecular weights anddiverse chemical types, mainly natural and synthetic polymers, such asglycosaminoglycans, hyaluronic acids, heparin, heparin sulfates,chondroitin sulfates, chondroitin-6-sulfates, chondroitin-4-sulfates,keratins, keratin sulfates, dermatins, dermatan sulfates, dextrans,plasma expanders, including polygeline (“Haemaccel”, degraded gelatinpolypeptide cross-linked via urea bridges, produced by “Behring”),“hydroxyethylstarch” (Hetastarch, HES) and extrans, food and drugadditives, soluble cellulose derivatives (e.g., methylcellulose,carboxymethylcellulose), polyaminoacids, hydrocarbon polymers (e.g.,polyethylene), polystyrenes, polyesters, polyamides, polyethylene oxides(e.g. polyethyleneglycols, polycarboxyethyleneglycols, polycarboxylatedpolyethyleneglycols), polyvinnylpyrrolidones, polysaccharides,polypyranoses, alginates, assimilable gums (e.g., xanthan gum),peptides, injectable blood proteins (e.g., serum albumin), cyclodextrin,and derivatives thereof.

In one embodiment, polysaccharides may be homo-polysaccharides, while inanother embodiment, they may be hetero-polysaccharides.

In one embodiment, examples of monomers, dimers, and oligomers which canbe employed as the conjugated moiety for producing Lipid-conjugates foruse in the methods of the invention may be mono- or disaccharides,trisaccharides, oligopeptides, carboxylic acids, dicarboxylic acids,fatty acids, dicarboxylic fatty acids, salicylates, salicyclic acids,acetyl salicylic acids, aspirins, lactobionic acids, maltoses, aminoacids, glycines, glutaric acids, succinic acids, dodecanoic acids,didodecanoic acids, bile acids, cholic acids, cholesterylhemisuccinates,and di- and trisaccharide unit monomers of polysaccharides,polypyranoses, and/or glycosaminoglycans including heparins, heparansulfates, hyaluronic acids, chondroitins, chondroitin sulfates,chondroitin-6-sulfates, chondroitin-4-sulfates, dermatins, dermatansulfates, keratins, keratan sulfates, or dextrans.

In some cases, according to embodiments of the invention, the monomer orpolymer chosen for preparation of the Lipid-conjugate may in itself haveselect biological properties. For example, both heparin and hyaluronicacid are materials with known physiological functions. In the presentinvention, however, the Lipid-conjugates formed from these substances asstarting materials display a new and wider set of pharmaceuticalactivities than predicted from the administration of either heparin orhyaluronic acid that are not covalently linked to a phospholipid. It canbe shown, by standard comparative experiments as described below and inU.S. application Ser. No. 10/952,496, incorporated herein by reference,that phosphatidylethanolamine (PE) linked to hyaluronic acid (CompoundXXII), to heparin (Compound XXIV), to chondroitin sulfate A (CompoundXXV), to carboxymethylcellulose (Compound XXVI), to Polygeline(haemaccel) (Compound XXVII), or to hydroxyethylstarch (CompoundXXVIII), are far superior in terms of potency and range of usefulpharmaceutical activity to the free conjugates (the polymers above andthe like). In fact, these latter substances are, in general, notconsidered useful in methods for treatment of most of the diseasesdescribed herein, including the treatment of dermatologic conditions.Thus, the combination of a phospholipid such asphosphatidylethanolamine, or related phospholipids which differ withregard to the polar head group, such as phosphatidylserine (PS),phosphatidylcholine (PC), phosphatidylinositol (PI), andphosphatidylglycerol (PG), results in the formation of a compound whichhas novel pharmacological properties when compared to the startingmaterials alone.

The biologically active Lipid-conjugates described herein can have awide range of molecular weights, e.g., above 50,000 (up to a few hundredthousands) when it is desirable to retain the Lipid conjugate in thevascular system and below 50,000 when targeting to extravascular systemsis desirable. The sole limitation on the molecular weight and thechemical structure of the conjugated moiety is that it does not resultin a Lipid-conjugate devoid of the desired biological activity, or leadto chemical or physiological instability to the extent that theLipid-conjugate is rendered useless as a drug in the method of usedescribed herein.

In one embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (A):

wherein

-   -   L is a lipid or a phospholipid;    -   Z is either nothing, ethanolamine, serine, inositol, choline,        phosphate, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer; and    -   n is a number from 1 to 1000;    -   wherein any bond between L, Z, Y and X is either an amide or an        esteric bond.

In one embodiment, L is phosphatidyl, Z is ethanolamine, wherein L and Zare chemically bonded resulting in phosphatidylethanolamine, Y isnothing, and X is carboxymethylcellulose. In another embodiment, L isphosphatidyl, Z is ethanolamine, wherein L and Z are chemically bondedresulting in phosphatidylethanolamine, Y is nothing, and X is dextran.In another embodiment, L is phosphatidyl, Z is ethanolamine, wherein Land Z are chemically bonded resulting in phosphatidylethanolamine, Y isnothing, and X is a glycosaminoglycan. In one embodiment, thephosphatidylethanolamine moiety is dipalmitoyl phosphatidylethanolamine.In another embodiment, the phosphatidylethanolamine moiety isdimyristoyl phosphatidylethanolamine.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (I):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms; and    -   X is either a physiologically acceptable monomer, dimer,        oligomer or a physiologically acceptable polymer; and    -   n is a number from 1 to 1,000;    -   wherein if Y is nothing the phosphatidylethanolamine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylethanolamine via an amide bond.

In one embodiment, compounds for use in the methods of the inventioncomprise one of the following as the conjugated moiety X: acetate,butyrate, glutarate, succinate, dodecanoate, didodecanoate, maltose,lactobionic acid, dextran, alginate, aspirin, cholate,cholesterylhemisuccinate, carboxymethyl-cellulose, heparin, hyaluronicacid, chondroitin sulfate, polygeline (haemaccel), polyethyleneglycol,polycarboxylated polyethylene glycol, a glycosaminoglycan, apolysaccharide, a hetero-polysaccharide, a homo-polysaccharide, or apolypyranose. The polymers used as starting material to prepare thePE-conjugates may vary in molecular weight from 1 to 2,000 kDa.

Examples of phosphatidylethanolamine (PE) moieties are analogues of thephospholipid in which the chain length of the two fatty acid groupsattached to the glycerol backbone of the phospholipid varies from 2-30carbon atoms length, and in which these fatty acids chains containsaturated and/or unsaturated carbon atoms. In lieu of fatty acid chains,alkyl chains attached directly or via an ether linkage to the glycerolbackbone of the phospholipid are included as analogues of PE. In oneembodiment, the PE moiety is dipalmitoyl-phosphatidyl-ethanolamine. Inanother embodiment, the PE moiety isdimyristoyl-phosphatidyl-ethanolamine.

Phosphatidyl-ethanolamine and its analogues may be from various sources,including natural, synthetic, and semisynthetic derivatives and theirisomers.

Phospholipids which can be employed in lieu of the PE moiety areN-methyl-PE derivatives and their analogues, linked through the aminogroup of the N-methyl-PE by a covalent bond; N,N-dimethyl-PE derivativesand their analogues linked through the amino group of theN,N-dimethyl-PE by a covalent bond, phosphatidylserine (PS) and itsanalogues, such as palmitoyl-stearoyl-PS, natural PS from varioussources, semisynthetic PSs, synthetic, natural and artifactual PSs andtheir isomers. Other phospholipids useful as conjugated moieties in thisinvention are phosphatidylcholine (PC), phosphatidylinositol (PI),phosphatidic acid and phosphoatidylglycerol (PG), as well as derivativesthereof comprising either phospholipids, lysophospholipids, phosphatidicacid, sphingomyelins, lysosphingomyelins, ceramide, and sphingosine.

For PE-conjugates and PS-conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through the nitrogen atom of thephospholipid polar head group, either directly or via a spacer group.For PC, PI, and PG conjugates, the phospholipid is linked to theconjugated monomer or polymer moiety through either the nitrogen or oneof the oxygen atoms of the polar head group, either directly or via aspacer group.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (II):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein if Y is nothing, the phosphatidylserine is directly        linked to X via an amide bond and if Y is a spacer, the spacer        is directly linked to X via an amide or an esteric bond and to        the phosphatidylserine via an amide bond.

In one embodiment, the phosphatidylserine may be bonded to Y, or to X ifY is nothing, via the COO⁻ moiety of the phosphatidylserine.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (III):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phosphatidyl, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (V):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, inositol, choline, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In one embodiment of the invention, phosphatidylcholine (PC),phosphatidylinositol (PI), phosphatidic acid (PA), wherein Z is nothing,and phosphatidylglycerol (PG) conjugates are herein defined as compoundsof the general formula (III).

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (VIII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXa):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (IXb):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the phospholipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (X):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer, or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the ceramide phosphoryl, Z, Y and X is        either an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XI):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein if Y is nothing the sphingosyl is directly linked to X        via an amide bond and if Y is a spacer, the spacer is directly        linked to X and to the sphingosyl via an amide bond and to X via        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, ethanolamine, serine, inositol, choline, or        glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the ceramide, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIII):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the diglyceryl, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIV):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XV):

wherein

-   -   R₁ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the glycerolipid, Z, Y and X is either        an amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is a linear, saturated, mono-unsaturated, or        poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XVIII):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XIX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XX):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

In another embodiment, the compound for use in the present invention isrepresented by the structure of the general formula (XXI):

wherein

-   -   R₁ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   R₂ is either hydrogen or a linear, saturated, mono-unsaturated,        or poly-unsaturated, alkyl chain ranging in length from 2 to 30        carbon atoms;    -   Z is either nothing, choline, phosphate, inositol, or glycerol;    -   Y is either nothing or a spacer group ranging in length from 2        to 30 atoms;    -   X is a physiologically acceptable monomer, dimer, oligomer or        polymer wherein X is a glycosaminoglycan; and    -   n is a number from 1 to 1000;    -   wherein any bond between the lipid, Z, Y and X is either an        amide or an esteric bond.

For any or all of the compounds represented by the structures of thegeneral formulae (A), (I), (II), (III), (IV), (V), (VI), (VII), (VIII),(IX), (IXa), (IXb), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI),(XVII), (XVIII), (XIX), (XX), (XXI), and (XXII) hereinabove: In oneembodiment, X is a glycosaminoglycan. According to this aspect and inone embodiment, the glycosaminoglycan may be, inter alia, hyaluronicacid, heparin, heparan sulfate, chondroitin sulfate, keratin, keratansulfate, dermatan sulfate or a derivative thereof. In anotherembodiment, X is not a glycosaminoglycan. In another embodiment, X is apolysaccharide, which in one embodiment is a hetero-polysaccharide, andin another embodiment, is a homo-polysaccharide. In another embodiment,X is a polypyranose.

In another embodiment, the glycosaminoglycan is a polymer ofdisaccharide units. In another embodiment, the number of thedisaccharide units in the polymer is m. In another embodiment, m is anumber from 2-10,000. In another embodiment, m is a number from 2-500.In another embodiment, m is a number from 2-1000. In another embodiment,m is a number from 50-500. In another embodiment, m is a number from2-2000. In another embodiment, m is a number from 500-2000. In anotherembodiment, m is a number from 1000-2000. In another embodiment, m is anumber from 2000-5000. In another embodiment, m is a number from3000-7000. In another embodiment, m is a number from 5000-10,000. Inanother embodiment, a disaccharide unit of a glycosaminoglycan may bebound to one lipid or phospholipid moiety. In another embodiment, eachdisaccharide unit of the glycosaminoglycan may be bound to zero or onelipid or phospholipid moieties. In another embodiment, the lipid orphospholipid moieties are bound to the —COOH group of the disaccharideunit. In another embodiment, the bond between the lipid or phospholipidmoiety and the disaccharide unit is an amide bond.

In another embodiment, the chondroitin sulfate may be, inter alia,chondroitin-6-sulfate, chondroitin-4-sulfate or a derivative thereof.

In one embodiment of the invention, Y is nothing. Non-limiting examplesof suitable divalent groups forming the optional bridging group (whichin one embodiment, is referred to as a spacer) Y, according toembodiments of the invention, are straight or branched chain alkylene,e.g., of 2 or more, preferably 4 to 30 carbon atoms, —CO-alkylene-CO,—NH-alkylene-NH—, —CO-alkylene-NH—, —NH-alkylene-NH, CO-alkylene-NH—, anamino acid, cycloalkylene, wherein alkylene in each instance, isstraight or branched chain and contains 2 or more, preferably 2 to 30atoms in the chain, —(—O—CH(CH₃)CH₂—)_(x)— wherein x is an integer of 1or more.

According to embodiments of the invention, in addition to thetraditional phospholipid structure, related derivatives for use in thisinvention are phospholipids modified at the C1 or C2 position to containan amine, ether or alkyl bond instead of an ester bond. In oneembodiment of the invention, the alkyl phospholipid derivatives andether phospholipid derivatives are exemplified herein.

In one embodiment of the invention, the sugar rings of theglycosaminoglycan are intact. According to this aspect and in oneembodiment, intact refers to closed. In another embodiment, intactrefers to natural. In another embodiment, intact refers to unbroken.

In one embodiment of the invention, the structure of the lipid orphospholipid in any compound according to the invention is intact. Inanother embodiment, the natural structure of the lipid or phospholipidsin any compound according to the invention is maintained.

In one embodiment, the compounds for use in the present invention arebiodegradable.

In one embodiment, the compound according to the invention isphosphatidylethanolamine bound to aspirin. In one embodiment, thecompound according to the invention is phosphatidylethanolamine bound toglutarate.

In some embodiments, the compounds for use are as listed in Table 1below.

TABLE 1 Phospholipid Spacer Polymer (m.w.) Compound PE None Hyaluronicacid XXII (2-2000 kDa) Dimyristoyl-PE None Hyaluronic acid XXIII PE NoneHeparin XXIV (0.5-110 kDa) PE None Chondroitin XXV sulfate A PE NoneCarboxymethyl- XXVI cellulose (20-500 kDa) PE Dicarboxylic PolygelineXXVII acid + (haemaccel) Diamine (4-40 kDa) PE None HydroxyethylstarchXXVIII PE Dicarboxylic Dextran XXIX acid + (1-2,000 kDa) Diamine PE NoneAspirin XXX PE Carboxyl amino Hyaluronic acid XXXI group (2-2000 kDa) PEDicarboxyl Hyaluronic acid XXXII group (2-2000 kDa) PE DipalmitoicHyaluronic acid XXXIII acid (2-2000 kDa) PE Carboxyl amino Heparin XXXIVgroup (0.5-110 kDa) PE Dicarboxyl Heparin XXXV group (0.5-110 kDa) PECarboxyl amino Chondroitin XXXVI group sulfate A PE DicarboxylChondroitin XXXVII group sulfate A PE Carboxyl amino Carboxymethyl-XXXVIII group cellulose (20-500 kDa) PE Dicarboxyl Carboxymethyl- XXXIXgroup cellulose (20-500 kDa) PE None Polygeline XL (haemaccel) (4-40kDa) PE Carboxyl amino Polygeline XLI group (haemaccel) (4-40 kDa) PEDicarboxyl Polygeline XLII group (haemaccel) (4-40 kDa) PE Carboxylamino Hydroxyethylstarch XLIII group PE Dicarboxyl HydroxyethylstarchXLIV group PE None Dextran XLV (1-2,000 kDa) PE Carboxyl amino DextranXLVI group (1-2,000 kDa) PE Dicarboxyl Dextran XLVII group (1-2,000 kDa)PE Carboxyl amino Aspirin XLVIII group PE Dicarboxyl Aspirin XLIX groupPE None Albumin L PE None Alginate LI (2-2000 kDa) PE None PolyaminoacidLII PE None Polyethylene glycol LIII PE None Lactobionic acid LIV PENone Acetylsalicylate LV PE None Cholesteryl- LVI hemmisuccinate PE NoneMaltose LVII PE None Cholic acid LVIII PE None Chondroitin sulfates LIXPE None Polycarboxylated LX polyethylene glycol Dipalmitoyl-PE NoneHyaluronic acid LXI Dipalmitoyl-PE None Heparin LXII Dipalmitoyl-PE NoneChondroitin LXIII sulfate A Dipalmitoyl-PE None Carboxymethyl- LXIVcellulose Dipalmitoyl-PE None Polygeline LXV (haemaccel) Dipalmitoyl-PENone Hydroxyethylstarch LXVI Dipalmitoyl-PE None Dextran LXVIIDipalmitoyl-PE None Aspirin LXVIII Dimyristoyl-PE None Heparin LXVIXDimyristoyl-PE None Chondroitin LXX sulfate A Dimyristoyl-PE NoneCarboxymethyl- LXXI cellulose Dimyristoyl-PE None Polygeline LXXII(haemaccel) Dimyristoyl-PE None Hydroxyethylstarch LXXIII Dimyristoyl-PENone Dextran LXXIV Dimyristoyl-PE None Aspirin LXXV PS None Hyaluronicacid LXXVI PS None Heparin LXXVII PS None Polygeline LXXVIII (haemaccel)PC None Hyaluronic acid LXXIX PC None Heparin LXXX PC None PolygelineLXXXI (haemaccel) PI None Hyaluronic acid LXXXII PI None Heparin LXXXIIIPI None Polygeline LXXXIV (haemaccel) PG None Hyaluronic acid LXXXV PGNone Heparin LXXXVI PG None Polygeline LXXXVII (haemaccel) PE NoneGlutaryl LXXXVIII

In one embodiment of the invention, the compounds for use in the presentinvention are any one or more of Compounds I-LXXXVIII. In anotherembodiment, the compounds for use in the present invention are CompoundXXII, Compound XXIII, Compound XXIV, Compound XXV, Compound XXVI,Compound XXVII, Compound XXVIII, Compound XXIX, Compound XXX, orpharmaceutically acceptable salts thereof, in combination with aphysiologically acceptable carrier or solvent. According to embodimentsof the invention, these polymers, when chosen as the conjugated moiety,may vary in molecular weights from 200 to 2,000,000 Daltons. In oneembodiment of the invention, the molecular weight of the polymer asreferred to herein is from 200 to 1000 Daltons. In another embodiment,the molecular weight of the polymer as referred to herein is from 200 to1000 Daltons. In another embodiment, the molecular weight of the polymeras referred to herein is from 1000 to 5000 Daltons. In anotherembodiment, the molecular weight of the polymer as referred to herein isfrom 5000 to 10,000 Daltons. In another embodiment, the molecular weightof the polymer as referred to herein is from 10,000 to 20,000 Daltons.In another embodiment, the molecular weight of the polymer as referredto herein is from 10,000 to 50,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 20,000 to70,000 Daltons. In another embodiment, the molecular weight of thepolymer as referred to herein is from 50,000 to 100,000 Daltons. Inanother embodiment, the molecular weight of the polymer as referred toherein is from 100,000 to 200,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 200,000 to500,000 Daltons. In another embodiment, the molecular weight of thepolymer as referred to herein is from 200,000 to 1,000,000 Daltons. Inanother embodiment, the molecular weight of the polymer as referred toherein is from 500,000 to 1,000,000 Daltons. In another embodiment, themolecular weight of the polymer as referred to herein is from 1,000,000to 2,000,000 Daltons. Various molecular weight species have been shownto have the desired biological efficacy.

In one embodiment of this invention, low molecular weightLipid-conjugates are defined hereinabove as the compounds of formula(I)-(XXI) wherein X is a mono- or disaccharide, carboxylateddisaccharide, mono- or dicarboxylic acids, a salicylate, salicylic acid,aspirin, lactobionic acid, maltose, an amino acid, glycine, acetic acid,butyric acid, dicarboxylic acid, glutaric acid, succinic acid, fattyacid, dodecanoic acid, didodecanoic acid, bile acid, cholic acid,cholesterylhemmisuccinate, a di- or tripeptide, an oligopeptide, atrisacharide, or a di- or trisaccharide monomer unit of heparin, heparansulfate, keratin, keratan sulfate, chondroitin, chondroitin-6-sulfate,chondroitin-4-sulfate, dermatin, dermatan sulfate, dextran, hyaluronicacid, glycosaminoglycan, or polypyranose.

Examples of suitable divalent groups forming the optional bridging groupY are straight- or branched-chain alkylene, e.g., of 2 or more,preferably 4 to 18 carbon atoms, —CO-alkylene-CO, —NH-alkylene-NH—,—CO-alkylene-NH—, cycloalkylene, wherein alkylene in each instance, isstraight or branched chain and contains 2 or more, preferably 2 to 18carbon atoms in the chain, —(—O—CH(CH₃)CH₂—)_(x)— wherein x is aninteger of 1 or more.

In another embodiment, in addition to the traditional phospholipidstructure, related derivatives for use in this invention arephospholipids modified at the C1 or C2 position to contain an ether oralkyl bond instead of an ester bond. These derivatives are exemplifiedhereinabove by the general formulae (VIII) and (IX).

In one embodiment of the invention, X is covalently conjugated to alipid. In another embodiment, X is covalently conjugated to a lipid viaan amide bond. In another embodiment, X is covalently conjugated to alipid via an esteric bond. In another embodiment, the lipid isphosphatidylethanolamine.

In one embodiment, cell surface GAGs play a key role in protecting cellsfrom diverse damaging agents and processes, such as reactive oxygenspecies and free radicals, endotoxins, cytokines, invasion promotingenzymes, and agents that induce and/or facilitate degradation ofextracellular matrix and basal membrane, cell invasiveness, white cellextravasation and infiltration, chemotaxis, and others. In addition,cell surface GAGs protect cells from bacterial, viral and parasiticinfection, and their stripping exposes the cell to interaction andsubsequent internalization of the microorganism. Enrichment of cellsurface GAGs would thus assist in protection of the cell from injuriousprocesses. Thus, in one embodiment of the invention, PLA2 inhibitors areconjugated to GAGs or GAG-mimicking molecules. In another embodiment,these Lipid-conjugates provide wide-range protection from diverseinjurious processes, and are effective in amelioration of diseases thatrequires cell protection from injurious biochemical mediators.

In another embodiment, a GAG-mimicking molecule may be, inter alia, anegatively charged molecule. In another embodiment, a GAG-mimickingmolecule may be, inter alia, a salicylate derivative. In anotherembodiment, a GAG-mimicking molecule may be, inter alia, a dicarboxylicacid.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from a dermalogic disorder,including a lipid or phospholipid moiety bonded to a physiologicallyacceptable monomer, dimer, oligomer, or polymer; and a pharmaceuticallyacceptable carrier or excipient.

In another embodiment, the invention provides a pharmaceuticalcomposition for treating a subject suffering from a dermalogic disorder,including any one of the compounds for use in the present invention orany combination thereof; and a pharmaceutically acceptable carrier orexcipient. In another embodiment, the compounds for use in the presentinvention include, inter alia, the compounds represented by thestructures of the general formulae as described hereinbelow: (A), (I),(II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (IXa), (IXb), (X),(XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII), (XIX), (XX),(XXI), (X), or any combination thereof.

Preparation of Compounds for use in the Present Invention

In one embodiment, the preparation of high molecular weightLipid-conjugates for use in the methods of the present invention is asdescribed in U.S. Pat. No. 5,064,817, which is incorporated fully hereinby reference. In one embodiment, these synthetic methods are applicableto the preparation of low molecular weight Lipid-conjugates as well,which in one embodiment are Lipid-conjugates comprising monomers anddimers as the conjugated moiety, with appropriate modifications in theprocedure as would be readily evident to one skilled in the art. Thepreparation of some low molecular weight Lipid-conjugates may beconducted using methods well known in the art or as described in U.S.patent application Ser. No. 10/952,496, which is incorporated herein byreference in its entirety.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following preferred specific embodiments are,therefore, to be construed as merely illustrative, and not limiting theremainder of the disclosure in any way whatsoever.

EXAMPLES

The abbreviations used in the examples below are:

-   PE=phosphatidyl-ethanolamine-   HA=hyaluronic acid-   Cpd=Compound-   Cpd XXII=dipalmitoyl-PE conjugated to HA-   Cpd XXIII=dimyristoyl-phosphatidyl-ethanolamine linked to HA-   Cpd XXIV=PE conjugated to heparin-   Cpd XXV=PE conjugated to chondroitin sulfate A (CSA)-   Cpd XXVI=PE conjugated to carboxymethyl cellulose (CMC)-   Cpd XXIX=PE conjugated to dextran-   Cpd XXX=PE conjugated to aspirin-   Cpd LXXXVIII=PE conjugated to glutaryl    The compounds used in the examples below were prepared as described    in U.S. patent application Ser. No. 10/952,496, which is fully    incorporated herein by reference.

Example 1 Skin Diseases, Contact Dermatitis and Psoriasis

Contact dermatitis is a widespread skin disease and is often attributedto a delayed type hypersensitivity response. Cutaneous, or skin,hypersensitivity reactions may occur in response to virtually anymaterial and may present clinically in either acute or chronic forms. Awidely-accepted system for invoking the delayed type hypersensitivityresponse is systemic sensitization to an antigen followed by its localapplication. Psoriasis is another common form of dermatitis marked byplaque-like formations, evident on extensor surfaces. As ahyperproliferative disorder of epithelial cells, drug therapies aretypically examined in cell cultures obtained from sufferers of thecondition.

Both secreted (sPLA) and cytosolic (cPLA) PLA2 have been identified inhuman skin. Their inflammatory roles have been determined in patientssuffering from inflammatory skin diseases such as psoriasis, althoughsome also play a role in maintaining healthy skin integrity.

Experiments 1.1-1.4 demonstrate that treatment of the animals afflictedwith a hypersensitivity reaction readily respond to the administrationof Lipid-conjugates, whether applied intraperitoneally (Table 1.1),subcutaneously (Table 1.2), or topically (Tables 1.3-1.4), as bothprophylactic and acute therapy.

Treatment Groups: In Experiments 1.1-1.2, “late sensitized only” micewere given topical application of oxazolone to both sides of one ear 24hours before measuring its swelling, while fully sensitized mice weretreated with topical application of oxazolone to their shaved stomachs,and then on day 6, with topical application of oxazolone to both sidesof one ear. In Experiments 1.3-1.4, treatment was as described above,except that oxazolone was applied to both sides of both ears.

Measurement of swelling: Swelling was measured in 0.1 mm increments bysubtracting the ear width of each individual mouse before treatment fromthe width after treatment. Percent inhibition was calculated by the netswelling of treated fully-sensitized ears (over that of the controlgroup A), divided by the net swelling of vehicle-treatedfully-sensitized ears.

In Experiment 1.1, mice were injected intraperitoneally daily from day 0until day 6 with vehicle (saline) or treatment compounds (40 mg CMC, 40mg Compound XXVI, or 5 mg betamethasone, in saline).

TABLE 1.1 Attenuation of Dermal Delayed Type Hypersensitivity Responseby Intraperitoneally Administered Lipid-Conjugate Swelling (ear widthafter Per- No. sensitization − cent of ear width before inhibi- GroupTreatment Mice sensitization)^(a) tion A Control 6 1.8 ± 1.0 — (latesensitized) B Fully sensitized + 6 18.5 ± 0.97 — saline C Fullysensitized + 6 19.8 ± 1.13 — carrier polymer (CMC) (0.4 μmol/kg) D Fullysensitized + 6  7.9 ± 1.37 66 Cpd XXVI (0.4 μmol/kg) E Fullysensitized + 6  6.5 ± 1.35 74 betamethasone (15 μmol/kg) ^(a)Data arepresented as mean ± S.D.

In Experiment 1.2, mice were injected subcutaneously adjacent to theoxazolone-challenged area, 3 h before application of oxazolone to theear and 1 h after application of oxazolone to the ear with vehicle(saline) or treatment compounds (40 mg CMC, 40 mg Compound XXVI, or 1 mgbetamethasone, in saline).

TABLE 1.2 Attenuation of Dermal Delayed Type Hypersensitivity Responseby Subcutaneously Adminstered Lipid-Conjugate Swelling (ear width afterPer- No. sensitization − cent of ear width before inhibi- GroupTreatment Mice sensitization) ^(a) tion A Control 5 4.1 ± 0.82 — (latesensitized) B Fully sensitized + 5 18.3 ± 0.82  — saline C Fullysensitized + 5 13.5 ± 2.17  35 carrier polymer (CMC) (0.4 μmol/kg) DFully sensitized + 5 5.9 ± 1.52 87 Cpd XXVI (0.4 μmol/kg) E Fullysensitized + 5 8.1 ± 1.19 72 betamethasone (3 μmol/kg) ^(a) Data arepresented as mean ± S.D.

In Experiment 1.3, mice were treated topically on both ears over thechallenged area daily beginning day 0 until day 6 with vehicle ortreatment compounds (20 μL of 0.1% Compound XXIX or dextran in 50% EtOHor 20 μl of Dermovat (steroid ointment)).

TABLE 1.3 Attenuation of Dermal Delayed Type Hypersensitivity Responseby Bilateral Topical Administration of Lipid Conjugate Swelling (earwidth after Per- No. sensitization − cent of ear width before inhibi-Group Treatment Mice sensitization) ^(a) tion A Control 5  1.5 ± 0.70 —(late sensitized only) B Fully sensitized + 5 24.3 ± 1.56 — saline CFully sensitized + 5 24.4 ± 2.4  — carrier polymer (dextran) (0.5μmol/kg) D Fully sensitized + 5 12.17 ± 1.52  53 Compound XXIX (0.5μmol/kg) E Fully sensitized + 5 10.6 ± 0.84 60 Dermovat (3 μmol/kg) ^(a)Data are presented as mean ± S.D.

In Experiment 1.4, mice were treated topically with vehicle or treatmentcompounds (20 μL of 0.1% Compound XXIX in 50% EtOH or 20 μl of Dermovat(steroid ointment)) on only one of the oxazolone-challenged ears 5 times4-6 hours following the oxazolone challenge.

TABLE 1.4 Attenuation of Dermal Delayed Type Hypersensitivity Responseby Unilateral Topical Administration of Lipid-Conjugate Swelling (earwidth after sensitization − Percent No. of ear width beforesensitization) ^(a) inhibition Group Treatment mice Left ear Both earsRight ear Left ear Right ear A Late sensitized only 10  1.0 ± 2.0 — — BFully sensitized +vehicle 10 23.0 ± 4.0 — — C Fully sensitized +Compound XXIX 7 20.0 ± 1.0 11.0 ± 1.0 14 46 (0.5 μmol/kg), on right earonly D Fully sensitized + Dermovat 7  7.0 ± 1.0  7.0 ± 1.0 63 63 (3μmol/kg) on right ear only ^(a) Data are presented as mean ± S.D.

As shown in Tables 1.1-1.4, treatment with the Lipid-conjugates reducedear swelling in mice showing a delayed-type hypersensitivity (DTH)response to oxazolone. The results presented in Table 1.4 demonstratethat although the topical administration of oxazolone was unilateral inboth cases, the steroid affected both ears, while the topically appliedLipid-conjugate affected only the area to which it was applied,indicating that the Lipid-conjugate acts locally rather thansystemically in this model, which may reduce potential side effects seenwith treatments that show systemic infiltration.

Experiment 1.5 demonstrates that Lipid-conjugates effectively inhibitthe proliferation of cultured psoriatic skin fibroblasts and Swiss 3T3cells. Fibroblasts of human psoriatic skin (dermis) cells, (fullcircles) or Swiss 3T3 cells (empty circles) were treated with CompoundXXVI at the indicated concentration for three days, after which thecells were counted (FIG. 1.1). The cell number of the control, untreatedgroup at the end of the three day incubation was taken as 100%. Forcomparison, carboxymethylcellulose was tested alone (data not shown).

Experiment 1.6: To show that Lipid-conjugates are effective in treatingpatients with contact dermatitis, a double-blind, placebo-controlledstudy was conducted in patients with contact dermatitis.

Drug preparation: A topical preparation of 1% Compound XXII (MW-50 kDa)was prepared by the Hadassah pharmacy (Jerusalem, Israel) using thefollowing w/w % ratios: Water 70.0, Cetyl Alcohol, 10.6, Paraffin, Whitesoft 10.6, Propylene Glycol 7.2, HyPE 1.0 and Sodium Dodecyl Sulfate0.6.

Study design and experimental procedures: The study group comprised atotal of 11 female patients aged 19-50 (mean age: 34.6). All thepatients had received a clinical diagnosis of contact dermatitis and apositive patch test to at least a single allergen. The diseasedistribution was symmetrical in all patients at the study sites. Allpatients exhibited contact dermatitis on the surface of their hands withsome patients exhibiting the disease on their forearms as well. Patientsrefrained from any systemic treatment for no less than a month beforethe initiation of the study and from topical treatment for no less thantwo weeks before the study. Disease severity was evaluated beforetreatment (day 0) and after a month of treatment (day 30) by thephysician assessment scoring criteria (ranging from 0 to 3) described inTable 1.5. In the initial analysis, the scores for each criteria wereassigned and summed to give a total score for each patient, with aminimum possible severity score of 0 and a maximum possible severityscore of 15. The range of severity scores for patients in the initialevaluation was 8-15.

TABLE 1.5 Physician assessment scoring criteria for contact dermatitisDryness 0-none 3-very dry Scaling 0-none 3-severe scaling Redness(erythema) 0-none 3-severe redness Pruritus (itching) 0-none 3-verypruritic Fissures 0-none 3-deep fissures

Each patient received two color-coded tubes of cream. One tube containedthe active pharmaceutical ingredient (Compound XXII), and the other tubecontained just the vehicle (placebo). Except for their label color, thetubes were identical in size, and the tube contents were identical incolor and odor. The doctor and the patients were unaware of which tubecontained the Compound XXII. Patients were instructed to consistentlyapply cream twice a day. They were instructed to apply the cream fromthe blue marked tube to their right hand and forearm and to apply creamfrom the pink marked tube to their left hand and forearm. The samedoctor evaluated the patients before and after treatment. The study wasapproved by the Helsinki Committee of Israeli Ministry of Health basedon animal safety data presented.

All 11 enrolled patients completed the one month study. The mean totalscore before treatment was 11.27±0.71. After one month of unilateraltreatment with the lipid conjugate, marked differences were visiblebetween the right and left hands and arms of the patients. On theCompound XXII-treated side, the average visual score was reduced by69.9%, while on the placebo-treated side, the average visual score wasreduced by 32.9% (p<0.005) (FIG. 1.2).

These experiments demonstrate that Lipid-conjugates are effectiveremedies for the management of various forms of dermatitis includingskin hypersensitivity reactions and contact dermatitis.

Example 2 Anti-Oxidant Therapy

There is evidence that allergic and inflammatory skin diseases likeatopic dermatitis, urticaria and psoriasis are mediated by oxidativestress, which damages cellular proteins, lipids, and DNA, leading tomembrane dysfunction and instability. Conversely, patients withdermatitis may be more susceptible to oxidative stress because ofdamaged skin may function as a weakened barrier. The skin isparticularly susceptible because it is exposed to endogenous andenvironmental pro-oxidant agents, leading to the harmful generation ofreactive oxygen species (ROS). The noxious effect of ROS, such asperoxide free radicals, on living tissue is known as oxidative stress ordamage.

In order to determine the effect of Lipid-conjugates on oxidative damageto proteins or cell membranes, tissue was exposed to hydrogen peroxide(H₂O₂) produced by (a) the enzyme glucose oxidase (GO) in the absence orpresence of additional membrane destabilizing agents such as PLA₂ or (b)by exposure to divalent cations, such as copper.

Experiments 2.1-2.3 demonstrate the ability of Lipid-conjugates topreserve cells from oxidative damage, as judged by the cells' retentionof both arachidonic acid and of low molecular weight intracellularsubstances.

Experiment 2.1: Confluent BGM (green monkey kidney epithelial) cellswere labeled with ³H-arachidonic acid. The cells were treated withCompound XXVI for 30 min prior to treatment with GO and PLA₂ (0.5 U/ml)(FIG. 2.1).

Experiment 2.2: BGM cells were labeled with ³⁵SO₄ overnight. The cellswere washed with DMEM (containing 10 mg/ml BSA) 4 times with PBS. Thecells were then incubated in DMEM supplemented with GO (an H₂O₂generator) for 90 min, and the culture medium was collected and countedfor ³⁵S radioactivity. For treatment with Compound XXVI, cells wereincubated with 3 or 10 μM Compound XXVI for 30 min prior to introductionof GO. Data are presented as mean±SEM for 5 replications. *p<0.005;**p<0.001 (FIG. 2.2).

Experiment 2.3 demonstrates the ability of Lipid-conjugates to inhibitthe oxidation of blood lipoprotein. Low density lipoprotein (LDL; 0.1μM) and or hydroperoxides (LOOH) were incubated in the absence andpresence of various concentrations of Compound XXII or HA at 37° C. Attime zero, 5 μM CuCl₂ was added to the dispersions, and the mixtureswere continuously monitored for oxidation products at 245 nm (FIG. 2.3).The absorbance at 245 (OD units) is depicted as a function of time.

These experiments demonstrate that administration of Lipid-conjugates isan effective therapy to prevent tissue damage induced by oxidativestress (associated with free radical and hydrogen peroxide production)by a plurality of mechanisms, including inhibiting the oxidation oflipoprotein, inhibiting arachidonic acid release, and preserving theintegrity of cell membranes (inhibiting GAG degradation), including redblood cell membranes, as described below. The efficacy ofLipid-conjugates in protecting against tissue damage induced byoxidative stress may contribute to their usefulness in preventing ortreating dermatitis.

Example 3 Hemolysis

Hemolysis, which in one embodiment, is the breakdown of red blood cells(RBC), may be either a primary disease in itself, or a syndromeassociated with another disease or physiological insult. In order todetermine the effect of Lipid-conjugates on hemolysis, red blood cellswere incubated in the presence of known membrane destabilizing agentsand the release of hemoglobulin into the extracellular medium wasdetected.

Experiment 3.1 demonstrates that the Lipid-conjugates serve to maintainthe stability of human red blood cells exposed to membrane-destroyingagents. Human RBC were washed in saline and suspended in Hanks buffer(pH 7.4). Hemolysis was induced in the absence or presence of 10 μMLipid-conjugates by treatment with either 5 U/ml streptolysin O (SLO),25 U/ml streptolysin S (SLS), or 5 μg/ml lysophosphatidylcholine(lyso-PC) for 20 min. The cell membranes were spun and the hemoglobincontent in the supernatant was determined by measuring the O.D. at 540nm (Table 3.1).

TABLE 3.1 Prevention of Hemolysis by Compound XXII, Compound XXVI andCompound XXIV HEMOLYSIS (O.D. AT 540 nm) Lipid-conjugate SLO SLS Lyso-PCNone 1.000 1.000 1.000 HA 1.000 1.000 1.875 Compound XXII-30* 0.6500.750 0.335 Compound XXII-60* 0.012 0.005 0.017 Compound XXII-110* 0.0050.002 0.012 Compound XXIV 0.002 1.100 0.002 Compound XXVI-60* 0.0120.005 0.002 Compound XXVI-110* 0.002 0.002 *The number expresses theamount of nmoles lipid conjugated to 1 mg of polymer.

These experiments demonstrate that the Lipid-conjugates are effectivetherapy in the treatment of cell membrane rupture and/or hemolysis.Thus, Lipid-conjugates protect against membrane destabilization, whichmay be a mechanism through which they are useful for the methods of thepresent invention. For example, Lipid-conjugates may protect againstmembrane destabilization that may lead to skin damage or dermatitis.

Example 4 PLA₂ Inhibition

The PLA₂ enzymes catalyze the hydrolysis of fatty acids attached tophospholipids on the plasma membrane. Arachidonic acid, the mainmetabolite released from these reactions, is a precursor for otherenzymatic reactions mediated by lipoxygenases and cyclooxygenases. Thesereactions produce prostaglandins and leukotrienes, which have a profoundeffect on inflammation in vivo. Therefore, PLA₂ inhibitors are capableof inhibiting inflammation via their ability to inhibit the productionof downstream inflammatory factors.

Experiment 4.1 was designed to determine the effect of Compound XXII,Compound XXV, Compound XXX, and Compound LXXXVIII on the inhibition ofthe Naja Naja Snake Venom PLA₂ enzyme in an in vitro fluorometric assay.The reaction of the PLA₂ enzyme and the PLA₂ enzyme substrate2-(6-(7-nitrobenz-2-oxa-1,3diazol-4-yl)amino)hexanoyl-1-hexadecanoyl-sn-glycero-3-phosphocholine(NHGP) yields a product, which can be detected using a fluorometer.Decreased absorbance indicates inhibition of the PLA₂ enzyme.

Methods

Compound XXII and Compound XXV were solubilized and diluted in D-PBS,and tested at final concentrations of 0.625, 0.125, 0.25, 0.5 and 1mg/ml. Compound XXX and Compound LXXXVIII were solubilized in 100%dimethyl sulfoxide (DMSO), diluted in D-PBS and tested at finalconcentrations of 0.01, 0.1 and 1 mg/ml. 1 mM NHGP was diluted in D-PBS,for a final concentration of 1 μM. The positive control, Mefenamic Acid(Sigma, M-4267), was tested at a final concentration of 0.1 mg/ml. ThePLA₂ enzyme is derived from the Naja Naja Snake Venom (Sigma, P6139) andtested at a final concentration of 5 Units/ml. The reaction was carriedout in 200 μl solution and initiated by addition of substrate.Fluorescence was read immediately and then every minute for 30 minutesfor a total of 30 readings. The fluorometer was set as follows:Excitation 450/50; Emission 530/25; Gain 50.

Results

Compound XXII inhibited the PLA₂ enzyme by 37%, 42%, 71% and 98% at0.125, 0.25, 0.5 and 1 mg/ml respectively (FIG. 4.1A) compared to 41%inhibition by 0.1 mg/ml mefenamic acid, which served as a positivecontrol. Compound XXV inhibited the PLA₂ enzyme, although with noapparent dose response, by 20%, 30% and 26% at 0.625, 0.125, 0.25 mg/ml(FIG. 4.1B). The inhibition of the PLA₂ enzyme by Compound LXXXVIII andCompound XXX could not be determined in this assay, due to difficultiesin solubilizing the compounds in DMSO, even after sonication.

Thus, Compound XXII inhibits the PLA₂ enzyme in a dose-dependent manner,indicating its ability to act as an anti-inflammatory drug. Otherexperiments showing anti-inflammatory effects of Lipid-conjugates aredemonstrated in U.S. application Ser. No. 10/952,496 filed Sep. 29, 2004and are hereby incorporated by reference.

Example 5 Toxicity Tests

Toxicity is a measure to the degree to which a compound or substance isdeleterious to an organism. Toxicological effects are generallydose-dependent. A therapeutic compound that is non-toxic, even at highdoses, would have an advantage over other compounds.

In Experiment 5.1, the Lipid-conjugates Compound XXII, Compound XXIV,Compound XXV and Compound XXVI were evaluated for toxicity. Toxicity wasevaluated in mice (3/group) one week after a single i.p. dose of 1000,500 or 200 mg/kg of Lipid-conjugates. Mortality rate, body weight, bloodcount (red and white cells), hematocrit, and internal organ histologyafter sacrifice were assessed. These parameters were compared inLipid-conjugate-treated and in control, untreated mice. Treatment withLipid-conjugates did not alter the parameters described above, with theexception of Compound XXIV, which induced hemorrhage.

Tables 5.1 and 5.2 depict the non-toxicity of Compound XXII asdemonstrated in acute (Table 5.1) and long-term (Table 5.2) toxicitytests.

TABLE 5.1 Results of acute (7 day) toxicity test - Compound XXII Dose ofCompound XXI (mg/kg Body weight (g) RBC × WBC × Hemat- body weight)Start Final 10⁶ 10³ ocrit % 0 (control) 21.9 ± 0.2 22.6 ± 0.3 10.7 ± 0.49.3 ± 0.3 45.0 ± 0.5 250 22.1 ± 0.4 23.1 ± 0.6 11.4 ± 0.1 7.7 ± 0.2 43.3± 0.7 500 21.4 ± 0.3 22.3 ± 0.4 11.5 ± 0.3 8.1 ± 1.3 44.7 ± 2.3 1000 21.7 ± 0.2 22.1 ± 0.2 10.9 ± 0.4 7.4 ± 0.6 40.3 ± 0.7RBC=red blood cells; WBC=white blood cells. Data are presented asmean±SEM.

For the long-term toxicity test, a group of 6 mice received an i.pinjection of 100 mg Lipid-conjugate (Compound XXII)/kg body weight 3times a week for 30 weeks (e.g., 180 mg total to a mouse weighing 20 g).Toxicity was evaluated as for Table 5.1. The results of the long-termtoxicity test are depicted in Table 5.2. There were no incidents ofmortality and no significant changes in body weight, red or white bloodcell count, or hematocrit induced by this treatment compared to control,untreated mice.

TABLE 5.2 Results of long-term (30 weeks) toxicity test - Compound XXIIDose of Compound XXII (mg/kg body Body weight, 3 times/ weight (g) RBC ×WBC × Hemat- week for 30 weeks) Final 10⁶ 10³ ocrit % 0 (control) 39.5 ±3.1 10.9 ± 0.8 9.3 ± 0.6 45.0 ± 0.8 100 39.0 ± 2.7 11.7 ± 0.7 8.1 ± 15 43.4 ± 4.9

In Experiment 5.2, Compounds XXII and XXV were evaluated for toxicityafter intravenous (i.v.) administration. Toxicity was evaluated inSprague-Dawley rats (5/group) five days after a four daily injectionsinto the tail of approximately 100 mg/kg of Lipid-conjugates or vehicle(PBS). Five days after the final injection, rats were anesthetized underisoflourane, the abdomen was opened, blood was collected by heartpuncture, urine was collected from the bladder, and organs werecollected into formalin. All animals were subjected to a full necropsyfollowing sacrifice, including examination of the external surface ofthe body, all orifices, and the cranial, thoracic and abdominal cavitiesand their contents.

All organs/tissues listed in Tables 5.3 and 5.4 were fixed and preservedin a 4% formaldehyde solution for at least a 48-hr fixation period. Inaddition, any other organs/tissues with gross macroscopic changes werepreserved as well in 4% formaldehyde solution. The preparation of slidesfor histopathology evaluation was performed by PAI, Maryland (Project03-1273). Histopathological changes were described and scored, usingsemiquantitative grading consisting of five grades (0-4) that reflectthe severity of the changes (0=unremarkable, 1=minimal, 2=mild,3=moderate, 4=marked).

Hematological, Biochemical and Urine Evaluation

Results demonstrated that neither Compound XXII nor Compound XXVindicated toxic effects on any of the systems or organs examined (Tables5.3, 5.4. and 5.5). All parameters were within the known normal rangefor this strain and age or were not significantly different from thecontrol group.

TABLE 5.3 Effect of i.v. Compound XXII and Compound XXV administrationon hematological parameters HEMATOLOGY Male Female Cpd XXII Cpd XXVControl Cpd XXII Cpd XXV Control Expected (n = 5) (n = 5) (n = 5) (n =5) (n = 5) (n = 5) Ranges WBC 7.68 ± 2.18 6.20 ± 1.74 6.70 ± 3.54 4.43 ±2.25  4.26 ± 0.77 5.62 ± 1.03  5-18 (×10E3/μL) RBC 7.68 ± 0.35 7.50 ±0.25 7.29 ± 0.33 7.47 ± 0.26  7.52 ± 0.48 7.51 ± 0.63  5-8.5 (×10E3/μL)HgB (g/dl) 15.28 ± 0.78  14.34 ± 0.51  14.22 ± 0.62  14.78 ± 0.61  14.12± 0.72 14.54 ± 1.29  10-16 HCT (%) 49.28 ± 2.85  47.58 ± 1.83  46.02 ±2.13  44.80 ± 2.23  45.14 ± 2.37 45.28 ± 4.35  35-50 MCV (fl) 64.16 ±2.28  63.42 ± 0.69  63.22 ± 3.22  59.94 ± 1.71  60.02 ± 1.37 60.40 ±2.86  50-75 MCH (pg) 19.88 ± 0.62  19.08 ± 0.25  19.50 ± 0.86  19.76 ±0.57  18.78 ± 0.29 19.36 ± 0.50  15-20 MCHC (g/dl) 30.98 ± 0.41  30.08 ±0.26  30.86 ± 0.43  32.96 ± 0.74  31.32 ± 0.51 32.08 ± 1.44  30-36 RDW(%) 15.40 ± 0.47  16.10 ± 0.67  15.84 ± 0.84  13.72 ± 0.54  13.76 ± 0.3414.14 ± 0.60  12-16 HDW 2.56 ± 0.06 2.57 ± 0.07 2.57 ± 0.08 2.60 ± 0.06 2.54 ± 0.10 2.59 ± 0.13 1.7-2.5 PLT 851.40 ± 154.66 927.20 ± 232.63877.0 ± 56.69 755.20 ± 166.63  910.6 ± 275.53 952.80 ± 125.66  900-1500(×10E3/μL) MPV (fl) 4.82 ± 0.53 4.82 ± 0.54 5.46 ± 0.32 4.86 ± 0.37 4.86 ± 0.18 5.34 ± 0.44  4-60 NEUT (%) 10.20 ± 1.30  15.80 ± 7.82 12.40 ± 5.41  15.00 ± 4.80  13.40 ± 5.68 9.80 ± 2.77 20-50 LYMP (%)87.20 ± 1.30  80.00 ± 7.38  84.20 ± 5.89  81.40 ± 4.34  82.40 ± 6.3586.60 ± 4.28   58-108 MONO (%) 2.40 ± 0.55 3.00 ± 1.22 3.20 ± 1.30 1.60± 1.34  2.80 ± 0.84 2.44 ± 1.89 0-8 EOS (%) 0.20 ± 0.45 1.20 ± 1.30 2.20± 4.92 1.80 ± 0.45  1.40 ± 1.34 0.80 ± 0.45 0-5 BASO (%) 0.00 ± 0.000.00 ± 0.00 0.00 ± 0.00 0.20 ± 0.45  0.00 ± 0.00 0.00 ± 0.00 0-1 Dataare presented as mean ± std. dev.

TABLE 5.4 Effect of i.v. Compound XXII and Compound XXV administrationon biochemical blood parameters BIOCHEMISTRY Male Female Cpd XXII CpdXXV Control Cpd XXII Cpd XXV Control Expected (n = 5) (n = 5) (n = 5) (n= 5) (n = 5) (n = 5) Ranges Glu (mg/dL)  164.60 ± 29.91 202.00 ± 83.30125.20 ± 22.95 124.40 ± 19.17 176.80 ± 35.58 176.60 ± 33.29 104.5-291.7Chol (mg/dL) 101.80 ± 6.76  95.00 ± 18.14  96.00 ± 11.38 90.80 ± 5.97105.80 ± 16.22  93.00 ± 15.81 30.46-107.8 UREA (mg/dL) 38.56 ± 4.8439.00 ± 4.95 41.80 ± 8.17 37.20 ± 4.97 33.80 ± 8.11 36.60 ± 5.5521.6-40.3 ALB (g/dL)  3.30 ± 0.10  3.46 ± 0.29  3.40 ± 0.12  3.54 ± 0.11 3.76 ± 0.23  3.70 ± 0.28 2.5-3.7 ALP (u/L) 292.80 ± 51.74 336.00 ±77.08 309.20 ± 74.45 202.00 ± 36.86 176.00 ± 20.33 194.80 ± 32.70 ALT(u/L) 47.80 ± 8.17 58.60 ± 7.44  55.00 ± 11.42 48.60 ± 6.91 50.00 ± 6.9648.60 ± 8.56  21.8-102.6 AST (u/L) 126.60 ± 11.97 110.60 ± 29.31 109.40± 19.72 123.40 ± 18.42 116.00 ± 49.20 91.00 ± 7.11  73.9-160.6 AMYL(u/L) 2722.60 ± 167.25 2599.20 ± 503.23 2774.00 ± 121.72 1787.20 ±106.83 1644.00 ± 182.62 1766.40 ± 240.51  1935.7-3271.88 Ca (mg/dL)10.76 ± 0.88  8.74 ± 2.12  8.82 ± 1.37  10.06 ± 0.92 10.66 ± 0.18 10.06± 1.11   6-10.6 CPK (u/L)  555.60 ± 185.54  410.00 ± 273.20  536.00 ±252.86  536.40 ± 191.27 308.00 ± 62.69 282.00 ± 84.59  339-1513 CREA(mg/dL)  0.53 ± 0.14  0.38 ± 0.09  0.37 ± 0.03  0.56 ± 0.04  0.49 ± 0.05 0.34 ± 0.15 0.34-0.57 Pi (mg/dL) 11.36 ± 1.49 10.21 ± 0.87 10.51 ± 1.1410.05 ± 0.62  9.58 ± 1.71  8.26 ± 0.92 5.89-8.27 TRIG (mg/dL) 110.20 ±41.70  82.00 ± 29.49 140.80 ± 44.57  65.60 ± 14.17  68.60 ± 29.36  42.60± 13.43 PROT (g/dL)  5.50 ± 0.14  5.84 ± 0.34  5.76 ± 0.38  5.60 ± 0.00 6.50 ± 0.25  6.30 ± 0.48 5.3-9.7 Globulins  2.20 ± 0.14  2.38 ± 0.24 2.36 ± 0.45  2.06 ± 0.11  2.74 ± 0.15  2.60 ± 0.31 2.09-6.8  (g/dL)Alb/Glob  1.48 ± 0.13  1.46 ± 0.21  1.50 ± 0.32  1.72 ± 0.15  1.38 ±0.13  1.44 ± 0.19 (ratio) T. BIL (mg/dL)  0.12 ± 0.04  0.13 ± 0.05  0.17± 0.05  0.10 ± 0.00  0.18 ± 0.04  0.14 ± 0.05 0.04-0.24 GGT (u/L)  0.00± 0.54  0.00 ± 0.00  0.00 ± 0.00  0.04 ± 0.09  0.00 ± 0.00  0.00 ± 0.00LDH (u/L)  24.29 ± 13.32  20.01 ± 18.70  20.41 ± 10.32  27.61 ± 21.4416.61 ± 4.43 13.68 ± 2.39 24.6-68.6 Cl (mmol/L) 99.60 ± 2.07 101.00 ±2.35  106.00 ± 4.53  101.80 ± 1.92  102.80 ± 3.11  107.20 ± 4.97  79.4-111.3 K (mmol/L)  6.24 ± 0.79  6.26 ± 0.78  6.08 ± 1.36  6.26 ±1.53  5.32 ± 0.38  5.80 ± 0.32 4-8 Na (mmol/L) 147.60 ± 1.14  142.00 ±1.58  137.60 ± 6.58  146.00 ± 2.92  140.20 ± 4.02  141.20 ± 1.79 135-155 Data are presented as mean ± std. dev.

TABLE 5.5 Effect of i.v. Compound XXII and Compound XXV administrationon urinalysis results URYNALYSIS Male Female Cpd XXII Cpd XXV ControlCpd XXII Cpd XXV Control (n = 5) (n = 5) (n = 5) (n = 5) (n = 5) (n = 5)Glu — — — — — — BIL — — — — — — Ketones — — — — — — S.G. 1.04 ± 0.011.03 ± 0.01 1.04 ± 0.01 1.04 ± 0.01 1.03 ± 0.01 1.04 ± 0.02 Eryth — — —— — — pH 6.60 ± 1.55 6.80 ± 0.45 6.80 ± 0.84 6.20 ± 0.45 7.00 ± 0.826.60 ± 0.55 T.S 6.88 ± 2.09 6.40 ± 2.12 8.22 ± 1.96 7.88 ± 4.00 6.13 ±1.66 7.80 ± 4.01 Leuk — — — — — — Data are presented as mean ± std. dev.

Macroscopic Evaluation

In addition, no macroscopic abnormalities were reported.

Histopathological Assessment

For Compound XXV, no treatment-related changes in histopathologicalassessment were noted (Table 5.6). For Compound XXII, treatment-relatedchanges were limited to the injection site (Table 5.7). These changeswere of minimal grade and consisted of subcutaneous mononuclear cellinfiltration and fibrosis. The mononuclear cell infiltration was notlimited to perivascular localization and therefore is not indicative ofimmunological reaction to the test compound. It is suggested that duringthe injection of the test compound, a minimal dose was deposited in thesubcutis, provoking a very minimal and limited inflammatory cellinfiltration, which should not be considered as an adverse reaction. Inaddition, in two animals, one male and one female, foreign body (hairshaft) minimal inflammation was noted. This change, although seen onlyin treated groups, is suggested to be caused by needle trauma and is notconsidered to be related to the test compound. Other observed changeshad comparable incidence in control and treated groups or are known tooccur frequently in untreated rats of the same age and strain, andtherefore are considered to be of no relation to the test compound. Suchchanges consisted of mononuclear cell infiltration in the liver andpresence of renal basophilic tubules.

TABLE 5.6 Effect of i.v. Compound XXV treatment on histopathology ofspecific organs. Compound XXV-treated Control, PBS-treated Males FemalesMales Females Brain 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Thymus 0 0 00 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Esophagus 0 0 0 0 0 0 0 0 M 0 0 0 0 00 0 0 0 0 0 Pancreas 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Liver 0 0 00 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Mononuclear cell 1 infiltrationPeriportal acute inflammation Spleen 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 0 Adrenal 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Lungs 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 0 0 Stomach 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 Heart 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Aorta 0 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 Kidneys 0 0 0 0 0 0 0 Cortex-basophilic 1 1 1 1 11 1 1 1 1 1 1 1 tubules Duodenum 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Jejunum 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Ileum 0 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 Colon 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Testes/Ovary 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Epididymides/Uterus0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Injection site (tail) 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 Subcutis - foreign 1 1 1 1 body (hair shaft)granulomatous inflammation Subcutis - Mononuclear 1 cell infiltrationSubcutis - Fibrosis Data are presented as mean ± std. dev.

TABLE 5.7 Effect of i.v. Compound XXII treatment on histopathology ofspecific organs. Compound XXII-treated Control, PBS-treated MalesFemales Males Females Brain 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Thymus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Esophagus 0 0 M 0 0 0 0 0M 0 0 0 0 0 0 0 0 0 0 0 Pancreas 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Liver 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Mononuclear cell 1 1infiltration Periportal acute inflammation Spleen 0 0 0 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 Adrenal 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Lungs0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Stomach 0 0 0 0 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 Heart 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Aorta 0 00 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Kidneys 0 0 0 0 0 0 0Cortex-basophilic 1 1 1 1 1 1 1 1 1 1 1 1 1 tubules Duodenum 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 0 0 0 Jejunum 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 0 Ileum 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Colon 0 0 0 0 0 0 0 00 0 0 0 0 0 0 0 0 0 0 0 Testes/Ovary 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 0 Epididymides/Uterus 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0Injection site (tail) 0 0 0 0 0 0 0 0 0 0 0 0 0 Subcutis - foreign 1 1body (hair shaft) granulomatous inflammation Subcutis - Mononuclear 1 11 1 1 1 1 cell infiltration Subcutis - Fibrosis 1 1 1 1

TABLE 5.8 Toxicity Test Results Dose (mg/injection) MK835 Survival (%)MK865 Survival (%) 0 100 100 0.6 100 100 2 66 100 6 33 66

Thus, the Lipid-conjugates have very low toxicity, as indicated in shortand long-term toxicity tests.

What is claimed is:
 1. A method of treating a dermatologic condition ina human subject, comprising the step of administering to said subject acomposition comprising a compound represented by the structure of thegeneral formula (I):

wherein R₁ is a linear, saturated, mono-unsaturated, orpoly-unsaturated, alkyl chain ranging in length from 2 to 30 carbonatoms; R₂ is a linear, saturated, mono-unsaturated, or poly-unsaturated,alkyl chain ranging in length from 2 to 30 carbon atoms; Y is nothing ora spacer group ranging in length from 2 to 30 atoms; X is apolysaccharide; and n is a number from 2 to
 1000. 2. The methodaccording to claim 1, wherein R₁ and R₂ are both palmitoyl moieties. 3.The method according to claim 1, wherein R₁ and R₂ are both myristoylmoieties.
 4. The method according to claim 1, wherein said dermatologiccondition is psoriasis.
 5. The method according to claim 1, wherein saiddermatologic condition is contact dermatitis.
 6. The method according toclaim 1, wherein said dermatologic condition is atopic dermatitis. 7.The method according to claim 1, wherein said polysaccharide isalginate.
 8. The method according to claim 1, wherein saidpolysaccharide is carboxymethylcellulose.
 9. The method according toclaim 1, wherein said polysaccharide is dextran.
 10. The methodaccording to claim 1, wherein n is a number from 2 to
 100. 11. Themethod according to claim 1, wherein the molecular weight of saidpolysaccharide is from 1-2,000 kDa.